AU779634B2

AU779634B2 - Diagnostics for the diagnosis or prognosis of glaucoma and related disorders

Info

Publication number: AU779634B2
Application number: AU31394/02A
Authority: AU
Inventors: Hua Chen; Pu Chen; Thai D Nguyen; Jon R Polansky
Original assignee: University of California Berkeley; University of California San Diego UCSD
Current assignee: University of California
Priority date: 1997-01-28
Filing date: 2002-04-03
Publication date: 2005-02-03
Anticipated expiration: 2018-01-09
Also published as: AU3139402A

Description

AUSTRALIA

Patents Act 1990 THE REGENTS OF THE UNIVERSITY OF CALIFORNIA COMPLETE SPECIFICATION STANDARD PATENT Invention Title: Diagnostics for the diagnosis or prognosis of glaucoma and related disorders The following statement is a full description of this invention including the best method of performing it known to us:a TITLE OF THE INVENTION: DIAGNOSTICS FOR THE DIAGNOSIS OR PROGNOSIS OF GLAUCOMA AND RELATED DISORDERS FIELD OF THE INVENTION: The present invention is in the fields of diagnostics, prognosis, and treatment, and concerns methods and reagents for diagnosing and treating glaucoma and related disorders.

BACKGROUND OF THE INVENTION: "Glaucomas" are a group of debilitating eye diseases that are the leading 15 cause of preventable blindness in the United States and other developed nations. Primary Open Angle Glaucoma ("POAG") is the most common form of glaucoma. The disease is characterized by the alteration of the trabecular meshwork, leading to obstruction of the normal ability of aqueous humor to leave the eye without closure of the space the "angle") between the iris 20 and cornea (see, Vaughan, D. et al., In: General Ophthalmology, Appleton Lange, Norwalk, Conn., pp. 213-230 (1992)). A characteristic of such obstruction in this disease is an increased intraocular pressure resulting in progressive visual loss and blindness if not treated appropriately and in a timely fashion.

The disease is estimated to affect between 0.4% and 3.3% of all adults over years old (Leske, M. C. et al., Amer. J. Epidemiol. 113:1843-1846 (1986); Bengtsson, Br. J. Ophthamol. 73:483-487 (1989); Strong, N. Ophthal.

Physiol. Opt. 12:3-7 (1992)). Moreover, the prevalence of the disease rises with age to over 6% of those 75 years or older (Strong, N. Ophthal. Physiol. Opt.

12:3-7 (1992)).

A link between the IOP response of patients to glucocorticoids and the disease of POAG has long been suspected. While only 5% of the normal population shows a high IOP increase (16 mm Hg) to topical glucocorticoid testing, greater than 40-50% of patients with POAG show this response. In addition, an Open Angle glaucoma may be induced by exposure to glucocorticoids. This observation has suggested that an increased or abnormal glucocorticoid response in trabecular cells may be involved in POAG (Zhan, G.L. et al., Exper. Eye Res. 54:211-218 (1992); Yun, AJ. et al., Invest. Ophthamol. Vis. Sci. 3(h2012-2022 (1989); Clark, Exper. Eye Res.

55:265 (1992); Klemetti, Acta Ophthamol. 68:29-33 (1990); Knepper, US.

Patent No. 4,617,299). The ability of glucocorticoids to induce a glaucoma-like condition has led to efforts to identify genes or gene products that would be induced by the cells of the trabecular meshwork in response to glucocorticoids (Polansky, J.R. et al., In: Glaucoma Update IV, Springer-Verlag, Berlin, pp. 20-29 (1991)). Initial efforts using short-term exposure to dexamethasone revealed only 15 changes in specific protein synthesis. Extended exposure to relatively high levels of dexamethasone was, however, found to induce the expression of related 66 kD and .55 kD proteins that could be visualized by gel electrophoresis (Polansky, J.R. et al., In Glaucoma Update IV, Springer-Verlag, Berlin, pp. 20-29 (1991)). The induction kinetics of these proteins as well as their dose response characteristics were similar to the kinetics that were required for steroid-induced IOP elevation in human subjects (Polansky, JR. et al., In: Glaucoma Update IV, Springer-Verlag, Berlin, pp. 29 (1991)). Problems of aggregation and apparent instability or loss of protein in the purification process were obstacles in obtaining a direct protein sequence.

Because increased IOP is a readily measurable characteristic of glaucoma, the diagnosis of the disease is largely screened for by measuring intraocular pressure (tonometry) (Strong, Ophthal. Physiol. Opt. 12:3-7 (1992), Greve, M. et al., Can. Ophthamol. 28:201-206 (1993)). Unfortunately, because glaucomatous and normal pressure ranges overlap, such methods are of limited value unless multiple readings are obtained (Hitchings, Br. J. OphthamoL 77-326 (1993); Tuck, M.W. et al., OphthaL Physiol. Opt. 13:227-232 (1993); Vaughan, D. et at, In: General Ophthamology, Appleton Lange, Norwalk, CT, pp. 213-230 (1992); Vernon, Eye 7:134-137 (1993)). For this reason, additional methods, such as direct examination of the optic disk and determination of the extent of a patient's visual field loss are often conducted to improve the accuracy of diagnosis (Greve, M. et al., Can. J. Ophthamol.

28201- 206 (1993)). Moreover, these techniques are of limited prognostic value.

04/08 '04 16:27 FAX 61 3 9639 2951 0007 3 Nguyen et al., U.S. Patent Application No: 08/649,432 filed May 17, 1996, the entire disclosure of which is hereby incorporated by reference as if set forth at length herein, disclosed a novel protein sequence highly induced by glucocorticoids in the endothellal lining cells of the human trabecular meshwork. Nguyen et U.S. Patent Application No: 08/649,432 also disclosed the cDNA sequence for that protein, the protein itself, molecules that bind to it, and nucleic acid molecules that encode it, and provided improved methods and reagents for diagnosing glaucoma and related disorders, as well as for diagnosing other diseases or conditions, such as cardiovascular, immunological, or other diseases or conditions that affect the expression or activity of the protein.

Any discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is solely for the purpose of S..providing a context for the present invention. It is not to be taken as an 15 admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed In Australia before the priority date of each claim of this application.

SUMMARY OF THE INVENTION One embodiment of the Invention is to provide a method for diagnosing I glaucoma in a patient which comprises the steps: incubating under conditions permitting nucleic acid hybridization: a marker nucleic acid molecule, said marker nucleic acid molecule comprising a nucleotide sequence of a polynucleotide that specifically hybridizes to a polynucleotide that is linked to 26 TIGR promoter, and a complementary nucleic acid molecule obtained from a Scell or a bodily fluid of said patient, wherein nucleic acid hybridization between said marker nucleic acid molecule and said complementary nucleic acid molecule obtained from said patient permits the detection of a polymorphism selected from the group consisting of TIGRmtl, TIGRmt2, TIGRmt3, TIGRmt4, TIGRmt5, and TIGRsv. whose presence is predictive of a mutation affecting TIGR response in said patient; permitting hybridization between said marker nucleic acid molecule and said complementary nucleic acid molecule obtained from said patient; and detecting the presence of said polymorphism selected from the group consisting of TIGRmtl, TIGRmt2, TIGRmt3, TIGRnt4, TIGRmt5, and TIGRsvl, whereln the detection of the polymorphism is diagnostic of glaucoma.

COMS ID No: SBMI-00856281 Received by IP Australia: Time 16:30 Date 2004-08-04 04/08 '04 16:27 FAX 61 3 9639 2951 1008 4 Another embodiment of the invention is to provide a method for prognosing glaucoma in a patient which comprises the steps: incubating under conditions permitting nucleic acid hybridization: a marker nucleic acid molecule, said marker nucleic acid molecule comprising a nucleotide sequence of a polynucleotide that specifically hybridizes to a nnlvynucnleot!de that is linkto a TIGR promoter, and a complementary nucleic acid molecule obtained from a cell or a bodily fluid of said patient, wherein nucleic acid hybridization between said marker nucleic acid molecule and said complementary nucleic acid molecule obtained from said patient permits the detection of a polymorphism selected from the group consisting of TIGRmt1, TIGRmt2, TIGRmt3, TIGRmt4, TIGRmt5, and TIGRsv1, whose presence is predictive of a mutation affecting TIGR response in said patient; permitting hybridization between said marker nucleic acid molecule and said complementary nucleic acid molecule obtained from said patient; and detecting the presence of *15 said polymorphism selected from the group consisting of TIGRmtl, TIGRmt2, TIGRmt3, TIGRmt4, TIGRmt5, and TIGRsvI, wherein the detection of the polymorphism is prognostic of glaucoma.

Another embodiment of the invention is to provide marker nucleic acid molecules capable of specifically detecting TIGRmtl, TIGRmt2, TIGRmt3, TIGRmt4, TIGRmtS, and TIGRsv1.

Another embodiment of the invention is to provide a method for diagnosing steroid sensitivity in a patient which comprises the steps: (A)

S

Incubating under conditions permitting nucleic acid hybridization: a marker nucleic acid molecule, the marker nucleic acid molecule comprising a 25 nucleotide sequence of a polynucleotide that specifically hybridizes to a polynucleotide that is linked to a TIGR promoter, and a complementary nucleic acid molecule obtained from a cell or a bodily fluid of a patient, wherein nucleic Sacid hybridization between the marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the patient permits the detection of a polymorphism selected from the group consisting of TIGRmtl, TIGRmt2, TIGRmt3, TIGRmt3, TIGRmt4, TIGRmt5, and TIGRsvl, whose presence is predictive of a mutation affecting TIGR response in the patient; (B) permitting hybridization between said marker nucleic acid molecule and the complementary nucleic acid molecule obtained from the patient; and (C) detecting the presence of the polymorphism selected from the group consisting COMS ID No: SBMI-00856281 Received by IP Australia: Time 16:30 Date 2004-08-04 04/08 '04 16:28 FAX 61 3 9639 2951 I1009 of TIGRml., TIGRmt2, TIGRmt3, TIGRmt4, TIGRmt5 and TIGRsvl, wherein the detection of the polymorphism is diagnostic of steroid sensitivity.

A further embodiment of the invention provides a diagnostic for the diagnosis or prognosis of glaucoma In a patient which comprises: a first marker nucleic acid molecule, said first marker nucleic acid molecule comprising a nucleotide sequence of a polynucleotide that specifically hybridizes to a polynucleotide that is linked to a TIGR promoter, and a complementary nucleic acid molecule obtained from a cell or bodily fluid of said patient, wherein nucleic acid hybridization between said first marker nucleic acid molecule and said complementary nucleic acid molecule obtained from said patient permits the detection of a polymorphism whose presence is 15 predictive of a mutation affecting TIGR response in said patient.

I: Preferably, the first marker nucleic acid is capable of specifically detecting nucleic acid selected from the group consisting of TIGRnmt, TIGRmt2, TIGRmt3, TIGRmt4, TIGRmt5 and TIGRsvl.

Preferably, the diagnostic also comprises a second marker nucleic acid molecule. Preferably, the second marker nucleic acid comprises a nucleotide sequence selected from the group consisting of SEQ ID NOS: 6-28 or a sequence complementary to any one of said SEQ ID NOS: 6-28, and more 25 preferably a sequence selected from the group consisting of SEQ ID NOS: 6-9, SEQ ID NOS: 12-13, SEQ ID NO:18 and SEQ ID NO: Preferably, the complementary nucleic acid is subjected to nucleic acid amplification, such as, for example, by means of polymerase chain reaction (PCR) or oligonucleotide ligation assay, to facilitate the detection of a single nucleotide polymorphism therein.

COMS ID No: SBMI-00856281 Received by IP Australia: Time 16:30 Date 2004-08-04 04/08 '04 16:28 FAX 61 3 9639 2951 i010 An additional embodiment of the present invention is to provide a method of treating glaucoma which comprises administering to a glaucomatous patient an effective amount of an agent that inhibits the synthesis of a TIGR protein.

Indeed, the molecules of the present invention may be used to diagnose s diseases or conditions which are characterised by alterations in the expression of extracellular proteins.

BRIEF DESCRIPTION OF THE FIGURES: Figures 1A, 1B, 1C, 1D and 1E provide the nucleic acid sequence of a TIGR promoter region (SEQ ID NO: 1) from an individual without glaucoma.

Figures 2A, 2B, 2C and 2D provide the location and sequence changes highlighted in bold associated with glaucoma mutants TIGRmtl, TIRGmt2, 15 TIGRmt3, TIGRmt4, TIGRmtS, and TIGRsvl (SEQ ID NO: 2).

Figures 3A, 3B, 3C, 3D, 3E, 3F and 3G provide nucleic acid sequences of a TIGR promoter, and TIGR exons, TIGR introns and TIGR downstream sequences (SEQ ID NO: 3, SEQ ID NO: 4, and SEQ ID NO: O Figure 4 provides a diagrammatic representation of the location of primers on the TIGR gene promoter for Single Strand Conformational Polymorphism (SSCP) analysis.

0- Figure 5 provides a diagrammatic representation of the TIGR exons and the arrangement of SSCP primers.

Figure 6 provides a homology analysis of TIGR homology with olfactomedin and olfactomedin-related proteins.

Figure 7 shows the nucleotide sequence of TIGR (SEQ ID NO: 26).

Figure 8 shows the amino acid sequence of TIGR (SEQ ID NO: 32).

DETAILED DESCRIPTION OF THE INVENTION: 1. Agents of the Invention COMS ID No: SBMI-00856281 Received by IP Australia: Time 16:30 Date 2004-08-04 As used herein, the term "glaucoma" has its art recognized meaning, and includes both primary glaucomas, secondary glaucomas, juvenile glaucomas, congenital glaucomas, and familial glaucomas, including, without limitation, pigmentary glaucoma, high tension glaucoma and low tension glaucoma and their related diseases. The methods of the present invention are particularly rel~vant to the diagnosis of POAG, QAG, juvenile glaucoma, and inherited glaucomas. The methods of the present invention are also particularly relevant to the prognosis of POAG, QAG, juvenile glaucoma, and inherited glaucomas. A disease or condition is said to be related to glaucoma if it possesses or exhibits a symptom of glaucoma, for example, an increased intra-ocular pressure resulting from aqueous outflow resistance (see, Vaughan, D. et al., In: General Ophthamology, Appleton Lange, Norwalk, CT, pp.21M-230 (1992)). The preferred agents of the present invention are discussed in detail below.

The agents of the present invention are capable of being used to diagnose the presence or severity of glaucoma and its related diseases in a patient suffering from glaucoma (a "glaucomatous patient"). The agents of the present invention are also capable of being used to prognose the presence or -severity of glaucoma and its related diseases in a person not yet suffering from any clinical manifestations of glaucoma. Such agents may be either naturally occurring or non-naturally occurring. As used herein, a naturally occurring molecule may be "substantially purified," if desired, such that one or more molecules that is or may be present in a naturally occurring preparation containing that molecule will have been removed or will be present at a lower concentration than that at whidch it would normally be found.

The agents of the present invention will preferably be "biologically active" with respect to either a structural attribute, such as the capacity of a nucleic acid to hybridize to another nucleic acid molecule, or the ability of a protein to be bound by antibody (or to compete with another molecule for such binding). Alternatively, such an attribute may be catalytic, and thus involve the capacity of the agent to mediate a chemical reaction or response.

As used herein, the term MfGR protein" refers to a protein having the amino acid sequence of SEQ ED NO: 32. As used herein, the agents of the present invention comprise nucleic acid molecules, proteins, and organic molecules.

As indicated above, the trabecular meshwork has been proposed to play an important role in the normal flow of the aqueous, and has been presumed to be the major site of outflow resistance in glaucomatous eyes. Human trabecular meshwork (HTM) cells are endothelial like cells which line the outflow channels by which aqueous humor exits the eye; altered synthetic function of the cells may be involved in the pathogenesis of steroid glaucoma and other types of glaucoma. Sustained steroid treatment of these cells are interesting because it showed that a major difference was observed when compared to 1-2 day glucocorticoid (GC) exposure.

This difference appears relevant to the clinical onset of steroid glaucoma (1-6 weeks).

Although trabecular meshwork cells had been found to induce specific proteins in response to glucocorticoids (see, Polansky, In: "Basic Aspects of Glaucoma Research III", Schattauer, New York 307-318 (1993)), efforts to purify the expressed protein were encumbered by insolubility and other problems. Nguyen, T.D. et al., (In: "Basic Aspects of Glaucoma Research III", Schattauer, New York, 331-343 (1993), herein incorporated by reference) used a molecular cloning approach to 15 isolate a highly induced mRNA species from glucocorticoid-induced human trabecular cells. .The mRNA exhibited a time course of induction that was similar to the glucocorticoid-induced proteins. The done was designated "IL2" (ATCC No: 97994, American Type Culture Collection, Rockville Maryland).

Nguyen et al., U.S. Patent Application No: 08/649,432 filed May 17, 1996, isolated a IL2 cone which encoded a novel secretory protein that is induced in cells of the trabecular meshwork upon exposure to glucocorticoids. It has been proposed t that this protein may become deposited in the extracellular spaces of the trabecular meshwork and bind to the surface of the endothelial cells that line the trabecular meshwork, thus causing a decrease in aqueous flow. Quantitative dot blot analysis and PCR evaluations have shown that the mRNA exhibits a progressive induction with time whereas other known GC-inductions from other systems and found in HTM cells (metallothionein, alpha-1 acid glycoprotein and alpha-1 antichymotrypsin) reached maximum level at one day or earlier. Of particular interest, the induction level of this clone was very high total cellular mRNA) with control levels undetectable without PCR method. Based on studies of 3 5

S

methionine cell labeling, the clone has the characteristics recently discovered for the major GC-induced extracellular glycoprotein in these cells, which is a sialenated, Nglycosylated molecule with a putative inositol phosphate anchor. The induction of mRNA approached 4% of the total cellular mRNA. The mRNA increased progressively over 10 days of dexamethasone treatment. The II. clone is 2.0 Kb whereas the Northern blotting shows a band of 2.5 Kb. Although not including a poly A tail, the 3' end of the done contains two consensus polyadenylation signals.

A genomic clone was isolated and designated PiTIGR clone (ATCC No: 97570, American Type Culture Collection, Rockville, Maryland). In-situ hybridization using the PiTIGR clone shows a TIGR gene and/or a sequence or sequences that specifically hybridize to the TIGR gene located at chromosome 1, q21-27, and more preferably to the TIGR gene located at chromosome 1, q22-26, and most preferably to the TIGR gene located at chromosome 1, q24. Clone PITIGR comprises human genomic sequences that specifically hybridize to the TIGR gene cloned into the BamHI site of vector pCYPAC (Ioannou et al., Nature Genetics, 6:84-89 (1994) herein incorporated by reference).

As used herein, the term "TIGR gene" refers to the region of DNA involved in producing a TIGR protein; it includes, without limitation, regions preceeding and following the coding region as well as intervening sequences between individual 15 coding regions.

As used herein, the term "TIGR exon" refers to any interrupted region of the TIGR gene that serves as a template for a mature TIGR mRNA molecule. As used herein, the term "TIGR intron" refers to a region of the TIGR gene which is noncoding and serves as a template for a TIGR mRNA molecule.

Localization studies using a Stanford G3 radiation hybrid panel mapped the S. TIGR gene near the D1S2536 marker with a LOD score of 6.0 (Richard et al., American Journal of Human Genetics 52.5: 915-921 (1993), herein incorporated by reference); Frazer et al., Genomics 14.3: 574-578 (1992), herein incorporated by reference; Research Genetics, Huntsville, Alabama). Other markers in this region include: D1S210; D1S1552; D1S2536; D1S2790; SHGC-12820; and D1S2558.

Sequences located upstream of the TIGR coding region are isolated and sequenced in a non-glaucomic individual. The upstream sequence is set forth in SEQ ID. No. 1. Sequence comparisons of the upstream region of a non-glaucoma individual and individuals with glaucoma identify a number of mutations in individuals with glaucoma. These mutations are illustrated in Figure 2. Five mutations are identified. TIGRmil is the result of a replacement of a cytosine with a guanine at position 4337 (SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3).

TIGRmt2 is the result of a replacement of a cytosine with a thymine at position 4950 (SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: TIGRmt3 is the result of an addition in the following order of a guanine, a thymine, a guanine, and a thymine (GTGT) at position 4998 (SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3).

TIGRmt4 is the result of a replacement of an adenine with a guanine at position 4256 (SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: TIGRmt5 is the result of a replacement of a guanine with an adenine at position 4262 (SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: One or more of TIGRmtl, TIGRmt2, TIGRmt3, TIGRmt4, and TIGRmt5 can be homozygous or heterozygous.

Sequence comparisons of the upstream region of a non-glaucoma individual and individuals with glaucoma identify at least one sequence variation in individuals with glaucoma. One such sequence variant is illustrated in Figure 2.

TIGRsvl is the result of a replacement of an adenine with a guanine at position 4406 (SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3).

Molecules comprising sequences upstream of the TIGR coding region provide useful markers for polymorphic studies. Such molecules include primers suitable for single strand conformational polymorphic studies, examples of which 15 are as follows: forward primer "Sk-la": 5'-TGA GGC TTC CTC TGG AAA C-3' (SEQ ID NO: reverse primer "ca2": 5'-TGA AAT CAG CAC ACC AGT AG-3' (SEQ ID N.O forward primer "CA2": 5'-GCA CCC ATA CCC CAA TAA TAG-3' (SEQ ID NO: reverse primer 5'-AGA GTT CCC CAG ATT TCA CC-3' (SEQ ID 2 NO: forward primer 5'-ATC TGG GGA ACT CTT CTC AG-3' (SEQ ID NO: 10); reverse primer "Pr+2(4A2)": 5'-TAC AGT TGT TGC AGA TAC G-3' (SEQ ID NO: 11); forward primer 5'-ACA ACG TAT CTG CAA CAA CTG-3' (SEQ ID NO: 12); reverse primer 5'-TCA GGC TA ACT GCA GAA CC-3' (SEQ ID NO: 13); forward primer 5'-TTG GTT CTG CAG TrA AGC C-3' (SEQ ID NO: 14); reverse primer "Pr+2(4A1)": 5'-AGC AGC ACA AGG GCA ATC C-3' (SEQ ID NO: 15); reverse primer 5'-ACA GGG CTA TAT TGT GGG-3' (SEQ ID NO: 16).

S In addition, molecules comprising sequences within TIGR exons provide useful markers for polymorphic studies. Such molecules include primers suitable for single strand conformational polymorphic studies, examples of which are as follows: forward primer "KSIX": 5'-CCT GAG ATG CCA GCT GTC C-3' (SEQ ID NO: 17); reverse primer "SK1XX": 5'-CTG AAG CAT TAG AAG CCA AC-3' (SEQ ID NO: 18); forward primer "KS2al": 5'-ACC TTG GAC CAG GCT GCC AG-3' (SEQ ID NO: 19); reverse primer "SK3" 5'-AGG IT GT CGA GTT CCA G-3' (SEQ ID NO: 20); forward primer "KS4": 5'-ACA ATT ACT GGC AAG TAT GG-3' (SEQ ID NO: 21); reverse primer "SK6A": 5'-CCT TCT CAG CCT TGC TAC C-3' (SEQ ID NO: 22); forward primer "KS5": 5'-ACA CCT CAG CAG ATG CTA CC-3' (SEQ ID NO: 23); reverse primer "SK8": 5'-ATG GAT GAC TGA CAT GGC C-3' (SEQ ID NO: 24); forward primer "KS6": 5'-AAG GAT GAA CAT GGT CAC C-3' (SEQ ID NO: The locations of primers: Sk-la, ca2, CA2, Pr+1, Pr-1, Pr+2(4A2), Pr-2(4A), Pr+3(4A), Pr-3(4A), Pr-3(4A), Pr+2(4A1), and Pr+1(4A) are diagramatically set forth in Figure 4. The location of primers: KS1X, SK1XX, Ks2al, SK3, KS4, SK6A, SK8, and KS6 are diagramatically set forth in Figure The primary structure of the TIGR coding region initiates from an ATG initiation site (SEQ ID NO:3, residues 5337-5339) and includes a 20 amino acid consensus signal sequence a second ATG (SEQ ID NO: 3, residues 5379-5381), indicating that the protein is a secretory protein. The nucleotide sequence for the TIGR coding region is depicted in Figure 7 (SEQ ID NO: 26). The protein contains an N-linked glycosylation site located in the most hydrophilic region of the 15 molecule. The amino terminal portion of the protein is highly polarized and adopts alpha helical structure as shown by its hydropathy profile and the Garier-Robison structure analysis. In contrast, the protein contains a 25 amino acid hydrophobic region near its carboxy terminus. This region may comprise a glucocorticoidinduced protein (GIP) anchoring sequence. The amino acid sequence of TIGR is depicted in Figure 8 (SEQ ID NO: 33).

Study of cyclohexamide treatment in the absence and presence of GC suggest that the induction of TIGR may involve factors in addition to the GC receptor. The TIGR gene may be involved in the cellular stress response since it is also induced by stimulants such as H202, 12-O-tetradecanolyphorbol-13-acetate (TPA), and high glucose; this fact may relate to glaucoma pathogenesis and treatment.

Sequence comparison of the upstream region identify a number of DNA motifs (cis elements). These DNA motifs or cis elements are shown in Figure 1.

These motifs include, without limitation, glucocorticoid response motif(s), shear stress response motif(s), NFcB recognition motif(s), and AP1 motif(s). The locations of these and other motifs are diagramatically set forth in Figure 1. As used herein, the term "cis elements capable of binding" refers to the ability of one or more of the described cis elements to specifically bind an agent Such binding may be by any chemical, physical or biological interaction between the cis element and the agent, including, but not limited, to any covalent, steric, agostic, electronic and ionic interaction between the cis element and the agent. As used herein, the term "specifically binds" refers to the ability of the agent to bind to a specified cis element but not to wholly unrelated nucleic acid sequences.

A preferred class of agents comprises TIGR nucleic acid molecules ("TIGR molecules"). Such molecules may be either DNA or RNA. A second preferred class of agents ("TIGR molecules") comprises the TIGR protein, its peptide fragments, fusion proteins, and analogs.

Expression of the rat PRL gene is highly restricted to pituitary lactotroph cells and is induced by the cAMI-dependent protein kinase A pathway. At least one of the redundant pituitary specific elements (PRL-FP111) of the proximal rat PRL promotor is required for this protein kinase A effect (Rajnarayan et aL, Molecular Endochronology 4: 502-512 (1995), herein incorporated by reference). A sequence corresponding to an upstream motif or cis element characteristic of PRL-FP111 is set forth in Figure 1 at residues 370-388 and 4491-4502, respectively. In accordance with the embodiments of the present invention, transcription of TIGR molecules can 15 be effected by agents capable of altering the biochemical properties or concentration of molecules that bind the PRL-FP111 upstream motif or cis element Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

A consensus sequence (GR/PR), recognized by both the glucocorticoid receptor of rat liver and the progesterone receptor from rabbit uterus, has been reported to be involved in glucocorticoid and progesterone-dependent gene expression (Von der Ahe et aL, Nature 313: 706-709 (1985), herein incorporated by reference). A sequence corresponding to a GC/PR upstream motif or cis element is set forth in Figure 1 at residues 433-445. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of glucocorticoid or progesterone or their homologues, including, but not limited to, the concentration of glucocorticoid or progesterone or their homologues bound to an GC/PR upstream motif or cis element. Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the sJudy of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

Shear stress motif (SSRE) or cis element has been identified in a number of genes including platelet-derived growth factor B chain, tissue plasnuinogen activator (tPA), ICAM-1 and TGF-J31 (Rsnick ei al., Proc. Nail. Acad. Sci. (USA) 80:.4591-4595 (1993), herein incorporated by reference). Transcription of these genes has been associated with humoral stimuli such as cytokines and bacterial products as well as hemodynamic stress forces. Sequences corresponding to a upstream shear stress motif or cis element are set forth in Figure 1 at residues 446-451, 1288-1293, 3597- 3602, 4771-4776, and 5240-5245, respectively. In accordance with the embodiments of the present invention, transcription of TIOR molecules can be effected by agents capable of altering the biochemical properties or concentration of molecules capable of binding the shear stress motif. Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

A consensus sequence for a glucocorticoid response upstream motif (GRE) or cis element has been characterized (Beato, Cell 56: 335-344 (1989); Becker ei al., Nature 324. 686-688 (1986), herein incorporated by reference; Sakai et al., Genes and Development 2: 1144-1154 (1988), herein incorporated by reference). Genes containing this upstream motif or cis element are regulated by glucocorticoids, progesterone, androgens and mineral corticoids (Beato, Cell 56.:335-344 (1989)).

Sequences corresponding to glucocorticoid response upstream motif or cis element are set forth in Figure 1 at residues 574-600, 1042-1056, 2444-2468, 2442-2269, 3536- 6 3563, 4574-4593, 4595-4614, 4851-4865, 4844-4864, 5079-5084, and 5083-5111, respectively. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of molecules capable of binding a glucocorticoid response upstream motif or cis element. Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

A sequence specific binding site (CBE) for the wild type nuclear phosphoprotein, p53, has been identified and appears to be associated with replication origins (Kern et al. Science 252: 1708-1711 (1991), herein incorporated by reference). A sequence corresponding to an CBE upstream motif or cis element is set forth in Figure 1 at residues 735-746. In accordance with the embodiments of the present invention, transcription of TGR molecules can be effected by agents capable of altering the biochemical properties or concentration of p53 or its homologues, including, but not limited to, the concentration of p53 or its homologues bound to an CBE upstream motif or cis element Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

Nuclear factor ets-like (NFE), a transcriptional activator that facilitates and c-Rel-dependent IgH 3' enhancer activity has been shown to bind to an NFE site in the Rel-dependent IgH 3' enhancer (Uinderson et al, European J. Immunology 27: 468-475 (1997), herein incorporated by reference). A sequence corresponding to an NFE upstream motif or cis element is set forth in Figure 1 at residues 774-795. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of nuclear factors or their homologues, including, but not limited to, the concentration of nuclear factors or their homologues bound to an NEE upstream motif or cis element. Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

upstream motif or cis element (KTF.1-CS) for a control element 3' to the human keratin 1 gene that regulates cell type and differentiation-specific expression has been identified (Huff et al., I. Biological Chemistry 268: 377-384 (1993), herein incorporated by reference). A sequence corresponding to an upstream motif or cis element characteristic of KTF.1-CS is set forth in Figure 1 at residues 843-854. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of KTF.1-CS or its homologues, including, but not limited to, the concentration of KTF.1-CS or its homologues bound to a KTF.1-CS upstream motif or cis element Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

A progesterone responsive element (PEE) that maps to the far upstream steroid dependent DNase hypersensitive site of chicken lysozyme chromatin has been characterized (Hecht et al., E.MBO 1.7:.2063-2073 (1988), herein incorporated by reference). The element confers hormonal regulation to a heterologous promoter and is composed of a cluster of progesterone receptor binding sites. A sequence corresponding to an upstream motif or cis element characteristic of PRE is set forth in Figure 1 at residues 987-1026. In accordance with the embodiments of the present invention, transcription of IIGR molecules can be effected by agents capable of altering the biochemnical properties or concentration of molecules capable of binding a progesterone responsive PEE upstream motif or cis element. Such agents may be useful in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

A sequence (ETF-EGFR) has been characterized which serves as a motif for a trans-active transcription factor that regulates expression of the epidermal growth factor receptor (Regec et al., Blood 85:2711-2719 (1995), herein incorporated by reference). A sequence corresponding to an ETF-EGFR upstream motif or cis element is set forth in Figure 1 at residues 1373-1388. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of nuclear factors or their homologues, including, but not limited to, the concentration of nuclear factors or their homologues bound to an ETF-EGFR upstream motif or cis element. Such agents can be used in the study of glaucoma pathogenesis. In 20 another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

A common trans-acting factor (SRE-cFos) has been shown to regulate skeletal and cardiac alpha-Actin gene transcription in muscle (Muscat et al., Molecular and Cellular Biology 10: 4120-4133 (1988), herein incorporated by reference). A sequence corresponding to an SRE-cFos upstream motif or cis element is se t forth in Figure 1 *at residues 1447-1456. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of nuclear factors or their homologues, including, but not limited to, the concentration of nuclear factors or their homologues bound to an SRE-cFos upstream motif or cis element Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

Alu repetitive elements are unique to primates and are interspersed within the human genome with an average spacing of 4Kb. While some Alu sequences are actively transcribed by polymerase Ml, normal transcripts may also contain Aluderived sequences in 5' or 3' untranslated regions (Jurka and Mikahanljaia, I. Mol.

Evolution 32: 105-121 (1991), herein incorporated by reference, Claveria and Makalowski, Nature 371: 751-752 (1994), herein incorporated by reference). A sequence corresponding to an Alu upstream motif or cis element is set forth in Figure 1 at residues 1331-1550. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of nuclear factors or their homologues, including, but not limited to, the concentration of nuclear factors or their homologues bound to an Alu upstream motif or cis element Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

A consensus sequence for a vitellogenin gene-binding protein (VBP) upstream motif or cis element has been characterized (Iyer et al., Molecular and Cellular Biology 11: 4863-4875 (1991), herein incorporated by reference). Expression of the VBP gene commences early in liver ontogeny and is not subject to circadian 20 control A sequence corresponding to an upstream motif or cis element capable of binding VBP is set forth in Figure 1 at residues 1786-1797. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of VBP or its homologues, including, but not limited to, the concentration of VBP or its S 25 homologues bound to an VBP upstream motif or cis element Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

A structural motif (Malt-CS) or cis element involved in the activation of all promoters of the maltose operons in Escherichia coli and Klebsiella pneumoniae has been characterized (Vidal-Ingigliardi et aL, I. MoL BioL 218: 323-334 (1991), herein incorporated by reference). A sequence corresponding to a upstream Malt-CS motif or cis element is set forth in Figure 1 at residues 1832-1841. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of molecules capable of binding the upstream Malt-CS motif or cis element. Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

A consensus sequence for an estrogen receptor upstream motif or cis element has been characterized (ERE) (Forman et al., Mol. Endocrinology 4: 1293-1301 (1990), herein incorporated by reference; de Verneuil et al., Nucleic Acid Res. 18: 4489-4497 (1990), herein incorporated by reference; Gaub et al, Cell 63: 1267-1276 (1990), herein incorporated by reference. A sequence corresponding to half an upstream motif or cis element capable of binding estrogen receptor is set forth in Figure 1 at residues 2166-2195, 3413-3429, and 3892-3896, respectively. In accordance with the embodiments of the present invention, transcription of TIOR molecules can be effected by agents capable of altering the biochemical properties or concentration, of the estrogen receptor or its homologues bound to an upstream motif or cis element.

Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

Certain protein-binding sites (NF-mutagen) in Ig gene enhancers which determine transcriptional activity and inducibility have been shown to interact with .20 nuclear factors (Lenardo et al, Science 236: 1573-1577 (1987), herein incorporated by reference). A sequence corresponding to an NF-mutagen upstream motif or cis element is set forth in Figure 1 at residues 2329-2338. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemnical properties or concentration of nuclear factors or their homologues, including, but not lim-ited to, the concentration of nuclear factors or their homologues bound to an NF-mutagen upstream motif or cis element. Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

A consensus sequence. for a transcriptional repressor of c-myc (myc-PRF) upstream motif or cis element has been identified (Kakkis et al., Nature 339: 718-719 (1989), herein incorporated by reference). Myc-PRF interacts with another widely distibuted protein, myc-CF1 (common factor which binds nearby and this association may be important in myc-PRF repression. A sequence corresponding to an upstream motif or cis element capable of binding myc-PRF is set forth in Figure 1 at residues 2403-24 16. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of myc-PEF or its homnologues, including, but not limited to, the concentration of myc-PRF or its homologues bound to an myc-PEF upstream motif or cis element Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

Human transcription factor activator protein 2 (AF2) is a transcription factor that has been shown to bind to Spi, nuclear factor 1 (NFl) and simian virus transplantation (SV40 T) antigen binding sites. It is developmentally regulated (Williams -and Tijan, Gene Dev. 5: 670-682 (1991), herein incorporated by reference; Mitchell et al., Genes Dev. 5: 105-119 (1991), herein incorporated by reference; Coutois et al, Nucleic Acid Research 18: 57-64 (1990), herein incorporated by reference; Comb et al., Nucleic Acid Research 18: 3975-3982 (1990), herein incorporated by reference; Winings et al., Nucleic Acid Research 19: 3709-3714 (1991), herein incorporated by reference). Sequences corresponding to an upstream motif or cis element capable of binding AF2 are set forth in Figure 1 at residues 2520-2535, and 5170-5187, respectively. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of AP2 or its homologues, including, but not limited to, the concentration of AP2 or its homiologues bound to an upstream motif or cis element. Such agents may be useful in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

Drosophila RNA polymerase UI heat shock transcription factor (HSTF) is a transcription factor that has been shown to be required for active transcription of an hsp 70 gene (Parker and Topol, Cell .37. 273-283 (1984), herein incorporated by reference). Sequences corresponding to an upstream motif or cis element capable of binding HSTF are set forth in Figure 1 at residues 2622-2635, and 5105-5132. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of I=T or its homologues, including, but not limited to, the concentration of HISTF or its homologues bound to an HSTF upstream motif or cis element. Such agents can be used in the study of glaucoma pathogenesis. in another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

A sequence corresponding to an upstream motif or cis element characteristic of SBF is set forth in Figure 1 at residues 2733-2743 (Shore et al., EMBO J. 6: 461-467 (1987), herein incorporated by reference). In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of molecules that bind the SBF upstream motif or cis element Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

An NFl motif or cis element has been identified which recognizes a family of at least six proteins (Courtois, et al., Nucleic Acid Res. 18: 57-64 (1990), herein incorporated by reference; Mul et al., J. Virol. 64: 5510-5518 (1990), herein incorporated by reference; Rossi et al, Cell 52. 405-414 (1988), herein incorporated by reference; Gounari et al., EMBO 1. 10: 559-566 (1990), herein incorporated by reference; Goyal et al., MoL Cell Bi. 10: 1041-1048 (1990); herein incorporated by reference; Mermond et al., Nature 331. 557-561 (1988), herein incorporated by reference; Gronostajski et al., Molecular and Cellular Biology 5: 964-971 (1985), herein incorporated by reference; Ilennighausen et al., EMBO 1. 5: 1367-1371 (1986), herein incorporated by reference; Chodosh et al., Cell 53: 11-24 (1988), herein incorporated by reference). The NFl protein will bind to an NFl motif or cis element either as a dimer (if the motif is palindromic) or as an single molecule (if the motif is not palindromic). The NFl protein is induced by TGFO (Faisst and Meyer, Nucleic Acid Research 20:-3-26 (1992), herein incorporated by reference). Sequences corresponding to an upstream motif or cis element capable of binding NFl are set forth in Figure 1 at residues 2923-2938, 4143-4167, and 4886-4900, respectively. In accordance with the embodiments of the present invention, transcription of IIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of NFl or its homologues, including, but not limited to, the concentration of NFl or its homiologues bound to an upstream motif or cis element Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

Conserved regulatory sequences (NF-MHCIIA/B) of a rabbit major histocompatability complex (MHC) class II gene are responsible for binding two distinct nuclear factors NF-MHCIIA and NF-MHCIIB and are believed to be involved in the regulation of coordinate expression of the class I genes eg. MHC class II gene in B lymphocytes (Sittisombut Molecular and Cellular Biology 5: 2034- 2041 (1988), herein incorporated by reference). A sequence corresponding to an NF- MHCIIA/B upstream motif or cis element is set forth in Figure I at residues 2936- 2944. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of NF-MHCIIA or NF-M-CIB or their homologues, including, but not limited to, the concentration of NF-MHCIIA or NF-MHCIIB or their homologues bound to an NF-MHCIIA/B upstream motif or cis element. Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

PEA 1 binding motifs or cis elements have been identified (Piette and Yaniv, EMBO J. 5:1331-1337 (1987), herein incorporated by reference). The PEA1 protein is 20 a transcription factor that is reported to bind to both the polyoma virus and c-fos enhancers A sequence corresponding to an upstream motif or cis element capable S. of binding PEA1 is set forth in Figure 1 at residues 3285-3298. In accordance with Sethe embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of '"25 PEA1 or its homologues, including, but not limited to, the concentration of PEA1 or its homologues bound to an upstream motif or cis element. Such agents can be used *in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

A conserved cis-acting regulatory element (ICS) has been shown to bind trans-acting constituitive nuclear factors present in lymphocytes and fibroblasts which are involved in the interferon (IFN)-mediated transcriptional enhancement of MHC class I and other genes (Shirayoshi t al., Proc. NatL Acad. Sci. (USA) 85: 5884- 5888 (1988), herein incorporated by reference). A sequence corresponding to an ICS upstream motif or cis element is set forth in Figure 1 at residues 3688-3699. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemnical properties or concentration of nuclear factors or their homologues, including, but not limited to, the concentration of nuclear factors or their homnologues bound to an ICS upstream motif or cis element. Such agents can be used in the study of glaucoma pathogenesis. in another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucomna.

A consensus sequence for an ISGF2 upstream motif or cis element has been characterized (Iman et al., Nucleic Acids Res. 18.:6573-6580 (1990), herein incorporated by reference; Harada et al, Cell 63: 303-312 (1990), herein incorporated by reference; Yu-Lee et al., Mol. Cell Bi. 10: 3087-3094 (1990), herein incorporated by reference; Pine et al., Mol. Cell Biol. 10: 32448-2457 (1990), herein incorporated by reference).

ISMF is induced by interferon a and y, prolactin and virus infections. A sequence corresponding to an upstream motif or cis element capable of binding ISGF2 is set forth in Figure 1 at residues 4170-4179. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of ISMF or its homologues, including, but not limited to, the concentration of ISGF2 or its homologues bound to 20 an upstream motif or cis element Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

A sequence corresponding, to an upstream motif or cis element capable of binding zinc is set forth in Figure 1 at residues 4285-4292. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of zinc. Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

A sequence corresponding to an upstream motif or cis element characteristic of CAP/ CRP-galO is set forth in Figure 1 at residues 4379-4404 (Tfaniguchi et al., Pwc. NatL Acad. Sci (USA) 76: 5090-5094 (1979), herein incorporated by reference). In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of molecules that bind the CAP/CRP-galO upstream motif or cis element. Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

Human iranscription factor activator protein 1 (AP1) is a transcription factor that has been shown to regulate genes which are highly expressed in transformed cells such as stromelysin, c-fos, a 1 -anti-trypsin and collagenase (Gutman and Wasylyk, EMBO 1. 9.7:2241-2246 (1990), herein incorporated by reference; Martin et aL, Proc. Natl. Acad. Sci. USA 85: 5839-5843 (1988), herein incorporated by reference; Jones et al., Genes and Development 2. 267-281 (1988), herein incorporated by reference; Faisst and Meyer, Nucleic Acid Research 20: 3-26 (1992), herein incorporated by reference; Kim et al., Molecular and Cellular Biology 10: 1492-1497 (1990), herein incorporated by reference: Baumhueter et al., EMBO J. 7: 2485-2493 (1988), herein incorporated by reference). The API transcription factor has been associated with genes that are activated by 12-O-tetradecanolyphorbol-13-acetate (TPA) (Gutman and Wasylyk, EMBO J.7: 2241-2246 (1990)). Sequences corresponding to an upstream motif or cis element capable of binding APi are set forth in Figure 1 at residues 4428-4434 and 4627-4639, respectively. In accordance with the embodiments of the present invention, transcription of TIGR molecules. can be effected by agents capable of altering the biochemical properties or concentration of API or its homologues, including, but not limited to, the concentration of AP1 or its homologues bound to an upstream motif or cis element. Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

*The sex-determining region of the Y chromosome gene, sry, is expressed in the fetal mouse for a brief period, just prior to testis differentiation. SRY is a DNA binding protein known to bind to a CACA-rich region in the sry gene (Vriz et al., Biochemistry and Molecular Biology International 37: 1137-1146 (1995), herein incorporated by reference). A sequence corresponding to an upstream motif or cis element capable of binding SRY is set forth in Figure 1 at residues 4625-4634. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of SRY or its homologues, including, but not limited to, the concentration of SRY or its homologues bound to an upstream motif or cis element.

Such agents may be useful in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

A sequence corresponding to an upstream motif or cis element characteristic of -C2GH s etfort in Fiue1 a&t_ reidues 4689-4711 (West et Molecular and Cellular Biology 7: 1193-1197 (1987), herein incorporated by reference). In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of GC2-GH or its homologues, including, but not limited to, the concentration of GC2-GH or its homologues bound to an upstream motif or cis element. Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

PEA 3 binding motifs or cis elements have been identified (Martin et al., Proc.

Nail. Acad. Sci. (USA) 85: 5839-5843 (1988), herein incorporated by reference; Gutman and Wasylyk, EMBO 1. 7: 2241-2246 (1990), herein incorporated by reference). The PEA3 protein is a transcription factor that is reported to interact with APi like proteins (Martin et al, Proc. NaiL Acad. Sci (USA) 85: 5839-584 (1988), herein incorporated by reference). Sequences corresponding to an upstream motif .:or cis element capable of binding PEA3 is set forth in Figure 1 at residues 4765-4769.

In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of PEA3 or its homologuzes, including, but not limited to, the concentration of PEA3 or its homologues bound to an upstream motif or cis element.

Mammalian interspersed repetitive (MIR) is an element -involved in the coding and processing sequences of mamla genes. The hM element is at least 260 bp in length and numbers about 1O copies within the mammalian genome (Murnane et al., Nucleic Acids Research 15: 2837-2839 (1995), herein incorporated by reference). A sequence corresponding to an MIR upstream motif or cis element is set forth in Figure 1 at residues 4759-4954. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of nuclear factors or their homologues, including, but not limited to, the concentration of nuclear factors or their homologues bound to an MIR upstream motif or cis element. Such agents can be used in the study of glaucoma pathogenesis. in another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used in the treatment of glaucoma.

Normal liver and differentiated hepatoma cell lines contain a hepatocytespecific nuclear factor (HNF-1) which binds cis-acting element sequences within the promoters of the alpha and beta chains of fibrinogen and alpha 1-antitrypsin (Baumhueter et al., EMBO 1. 8: 2485-2493, herein incorporated by reference). A sequence corresponding to an HNF-1 upstream motif or cis element is set forth in Figure 1 at residues 4923-4941. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of HNF-1 or its homologues, including, but not limited to, the concentration of HNF-1 or its homologues bound to an HNFL1 upstream motif or cis element Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In another embodiment such agents can be used 20 in the treatment of glaucoma.

A number of cis elements or upstream motifs have been associated with gene :regulation by steroid and thyroid hormones glucocorticoid. and estrogen)(Beato, Cell 56.:335-344 (1989), herein incorporated by reference; Brent et al., Mokeclar Endocrinology 89:1996-2000 (1989), herein incorporated by reference; Glass et al, Cell 54: 313-323 (1988), herein incorporated by reference; Evans, Science 240: 889-895 (1988), herein incorporated by reference).

A consensus sequence for a thyroid receptor upstream motif or cis element (TEE) has been Characterized (Beato, Cell 56.:335-344 (1989), herein incorporated by reference). A sequence corresponding to a thyroid receptor upstream motif or cis element is set forth in Figure 1 at residues 5151-5156. Thyroid hormones are capable of regulating genes containing a thyroid receptor upstream motif or cis element (Glass et al., Cell 54: 313-323 (1988), herein incorporated by reference).

Thyroid hormones can negatively regulate TIGR. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of molecules capable of binding a thyroid receptor upstream motif or cis element.

NFKB is a transcription factor that is reportedly associated with a number of biological processes including T-cell activation and cytokine regulation (Lenardo et aL, Cell 58:227-229 (1989), herein incorporated by reference). A consensus upstream motif or cis element capable of binding NFcB has been reported (Lenardo et al., Cell 58 227-229 (1989)). Sequences corresponding to an upstream motif or cis element capable of binding NFicB are set forth in Figure 1 at residues 5166-5175. In accordance with the embodiments of the present invention, transcription of TIGR molecules can be effected by agents capable of altering the biochemical properties or concentration of NFicB or its homologues, including, but not limited to, the concentration of NFcB or its homologues bound to an upstream motif or cis element.

Such agents can be used in the study of glaucoma pathogenesis. In another embodiment, such agents can also be used in the study of glaucoma prognosis. In Sanother embodiment such agents can be used in the treatment of glaucoma.

.Where one or more of the agents is a nucleic add molecule, such nucleic acid molecule may be sense, antisense or triplex oligonucleotides corresponding to any part of the TIGR promoter, TIGR cDNA, TIGR intron, TIGR exon or TIGR gene.

The TIGR promoter, or fragment thereof, of the present invention may be cloned into a suitable vector and utilized to promote the expression of a marker gene firefly luciferase (de Wet, Mol. Cell Biol. 7:725-737 (1987), herein incorporated by reference) or GUS (Jefferson et al., EMBO J. 6: 3901-3907 (1987), herein incorporated by reference)). In another embodiment of the present invention, a TIGR promoter may be cloned into a suitable vector and utilized to promote the expression of a TIGR gene in a suitable eukaryotic or prokaryotic host cell human trabecular cell, chinese hamster cell, E. coli). In another embodiment of the present invention, a TIGR promoter may be cloned into a suitable vector and utilized to promote the expression of a homologous or heterologous gene in a suitable eukaryotic or prokaryotic host cells human trabecular cell lines, chinese hamster cells, E. colt).

Practitioners are familiar with the standard resource materials which describe specific conditions and procedures for the construction, manipulation and isolation of macromolecules DNA molecules, plasmids, etc.), generation of recombinant organisms and the screening and isolating of clones, (see for example, Sambrook et al., In Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press (1989), herein incorporated by reference in its entirety; Old and Primrose, In Principles of Gene Manipulation: An Introduction To Genetic Engineering, Blackwell (1994), herein incorporated by reference).

The TIGR promoter or any portion thereof of the present invention may be used in a gel-retardation or band shift assay (Old and Primrose, In Principles of Gene Manipulation: An Introduction To Genetic Engineering, Blackwell (1994)).

Any of the cis elements identified in the present invention may be used in a gelretardation or band shift assay to isolate proteins capable of binding the cis element.

Suitable DNA fragments or molecules comprise or consist of -one or more of the following: sequences corresponding to an upstream motif or cis element characteristic of PRL-FP111 as set forth in Figure 1 at residues 370-388, and 4491- 4502, respectively, a sequence corresponding to an upstream motif or cis element capable of binding GR/PR as set forth in Figure 1 at residues 433-445, sequences corresponding to an upstream shear stress motif or cis element as set forth in Figure 1 at residues 446-451, 1288-1293, 3597-3602, 4771-4776, and 5240-5245, respectively, sequences corresponding to glucocorticoid response upstream motif or cis element .~as set forth in Figure 1 at residues 574-600, 1042-1056, 2444-2468, 2442-2269, 3536- 20 3563,4574-4593,4595-4614,4851-4865,4844-4864,5079-5084, 5083-5111, respectively, a sequence corresponding to an upstream motif or cis element capable of binding CBE as set forth in Figure 1 at residues 735-746, a sequence corresponding to an upstream motif or cis element capable of binding NFE as set forth in Figure 1 at residues 774-795, a sequence corresponding to an upstream motif or cis element capable of binding KTF.1-CS as set forth in Figure 1 at residues 843-854, a sequence corresponding to an upstream motif or cis element capable of binding PRE is set forth in Figure 1 at residues 987-1026, a sequence corresponding to an upstream motif or cis element capable of binding ETF-EGFR as set forth in Figure 1 at residues 1373-1388, a sequence corresponding to an upstream motif or cis element capable of binding SRE-cFos as set forth in Figure 1 at residues 1447-1456, a sequence corresponding to an upstream motif or cis element capable of binding Alu as set forth in Figure 1 at residues 1331-1550, a sequence corresponding to an upstne~m motif or cis element capable of binding VBP as set forth in Figure 1 at residues 1786-1797, a sequence corresponding to an upstream motif or cis element 35 capable of binding Malt-CS as set forth in Figure 1 at residues 1832-1841, sequences corresponding to an upstream motif or cis element capable of binding ERE as set forth in Figure 1 at residues 2167-2195, 3413-3429, and .3892-3896, respectively, a sequence corresponding to an upstream motif -or cis element capable of binding NFmutagen as set forth in Figure 1 at residues 2329-2338, a sequence corresponding to an upstream motif or cis element capable of binding myc-PRF as set forth in Figure I at4 residues.. '03A 2'J "C A iM sequences corresponding to an upstream motif or cis element capable of binding AP2 as set forth in Figure 1 at residues 2520-2535 and 5170-5187, respectively, sequences corresponding to an upstream motif or cis element capable of binding HSTF as set forth in Figure 1 at residues 2622-2635, and 5105-5132, respectively, a sequence corresponding to an upstream motif or cis element characteristic of SBF as set forth in Figure 1 at residues 2733-2743, sequences corresponding to an upstream motif or cis element capable of binding NF-1 as set forth in Figure 1 at residues 2923-2938, 4144-4157, and 4887-4900, respectively, a sequence corresponding to an upstream motif or cis element capable of binding NF- MHCIIA/B as set forth in Figure 1 at residues 2936-2944, a sequence corresponding to an upstream motif or cis element capable of binding PEAl as set forth in Figure 1 at residues 3285-3298, a sequence corresponding to an upstream motif or cis element capable of binding ICS as set forth in Figure 1 at residues 3688-3699, a sequence corresponding to an upstream motif or cis element capable of binding ISGF as set forth in Figure 1 at residues 4170-4179, a sequence corresponding to an upstream motif or cis element capable of binding zinc as set forth in Figure 1 at residues 4285-4293, a sequence corresponding to an upstream motif or cis element characteristic of CAP/CRP-galO as set forth in Figure 1 at residues 4379-4404, sequences corresponding to an upstream motif or cis element capable of binding AP1 as set forth in Figure 1 at residues 4428-4434, and 4627-4639, respectively, a sequence corresponding to an upstream motif or cis element capable of binding SRY as set forth in Figure 1 at residues 4625-4634, a sequence corresponding to an upstream motif or cis element characteristic of GC2 as set forth in Figure 1 at residues 4678-4711, a sequence corresponding to an upstream motif or cis element capable of binding PEA3 as set forth in Figure 1 at residues 4765-4769, a sequence corresponding to an upstream motif or cis element capable of MMR as set forth in Figure 1 at residues 4759-4954, a sequence corresponding to an upstream motif or cis element capable of binding NF-HNF-l as set forth in Figure 1 at residues 4923-4941, a sequence corresponding to a thyroid receptor upstream motif or cis element as set forth in Figure 1 at residues 5151-5156, and a sequence corresponding to an upstream motif or cis element capable of binding NFkB as set forth in Figure 1 at residues 5166-5175.

A preferred class of agents of the present invention comprises nucleic acid molecules will encode all or a fragment of "TIGR promoter" or flanking gene sequences. As used herein, the terms "TIGR promoter" or "promoter" is used in an expansive sense to refer to the regulatory sequence(s) that control mRNA production. Such sequences include RNA polymerase binding sites, glucocorticoid response elements, enhancers, etc. All such TIGR molecules may be used to diagnose the presence of glaucoma and severity of glaucoma. Such molecules may be either DNA or RNA.

Fragment nucleic acid molecules may encode significant portion(s) of, or indeed most of, SEQ ID NO: 1 or SEQ ID N. 3 or SEQ ID NO: 4 or SEQ ID NO: Alternatively, the fragments may comprise smaller oligonucleotides (having from about 15 to about 250 nucleotide residues, and more preferably, about 15 to about nucleotide residues.). Such oligonucleotides include SEQ ID NO. 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO. 14, SEQ ED NO. 15, SEQ ID NO. 16, SEQ ID NO, 17, SEQ ID NO .N 18, SEQ N. 19, SEQ ID NO: 20, SEQIED N.21, SEQ ID NO: 22, SEQ ID Na.23, SEQ ID NO: 24, SEQ ID Alternatively such oligonucleotides may derive from either the TIGR promoter, TIGR introns, TIGR exons, TIGR cDNA and TIGR downstream sequences comprise or consist of one or more of the following- sequences corresponding to an upstream motif or cis element characteristic of PRL-FP111 as set forth in Figure 1 at residues 370-388, and 4491-4502, respectively, a sequence corresponding to an 25 upstream motif or cis element capable of binding GR/PR as set forth in Figure 1 at residues 433-445, sequences corresponding to an upstream shear stress motif or cis element as set forth in Figure 1 at residues 446-451, 1288-1293, 3597-3602, 4771-4776, and 5240-5245, respectively, sequences corresponding to glucocorticoid response upstream motif or cis element as set forth in Figure 1 at residues 574-600,1042-1056, 2444-2468, 2442-2269, 3536-3563, 4574-4593, 45954614, 4851-4865, 4844-4864, 5079- 5084, 5083-5111, respectively, a sequence corresponding to an upstream motif or cis element capable of binding CBE as set forth in Figure 1 at residues 735-746, a sequence corresponding to an upstream motif or cis element capable of binding NFE as set forth in Figure 1 at residues 774-795, a sequence corresponding to an upstream motif or cis element capable of binding KTF.1-CS as set forth in Figure I at residues 843-854, a sequence corresponding to an upstream motif or cis element capable of binding PRE is set forth in Figure 1 at residues 987-1026, a sequence corresponding to an upstream motif or cis element capable of binding ErF-EGFR as set forth in Figure 1 at residues 1373-1388, a sequence corresponding to an upstream motif or cis element capable of binding SRE-cFos as set forth in Figure 1 at residues I 447-15, A i seqec correspon"dinto an upstreamn motif or cis element capable of binding Alu as set forth in Figure 1 at residues 1331-15,50, a sequence corresponding to an upstream motif or cis element capable of binding VBP as set forth in Figure 1 at residues 1786-1797, a sequence corresponding to an upstream motif or cis element capable of binding Malt-CS as set forth in Figure 1 at residues 1832-1841, sequences corresponding to an upstream motif or cis element capable of binding ERE as set forth in Figure 1 at residues 2167-2195, 3413-3429, and 3892-3896, respectively, a sequence corresponding to an upstream motif or cis element capable of binding NF-mutagen as set forth in Figure 1 at residues 2329-2338, a sequence corresponding to an upstream motif or cis element capable of binding myc-PEF as set forth in Figure 1 at residues 2403-2416, sequences corresponding to an upstream motif or cis element capable of binding AP2 as set forth in Figure 1 at residues 2520- 2535 and 5170-5187, respectively, sequences corresponding to an upstream motif or cis element capable of binding HSTF as set forth in Figure 1 at residues 2622-2635, and 5105-5132, respectively, a sequence corresponding to an upstream motif or cis element characteristic of SBF as set forth in Figure 1 at residues 2733-2743, sequences corresponding to an upstream motif or cis element capable of binding NF-1 as set forth in Figure 1 at residues 2923-2938, 4144-4157, and 4887-4900, respectively, a sequence corresponding to an upstream motif or cis element capable of binding NF- 25 MHCIIA/B as set forth in Figure 1 at residues 2936-2944, a sequence corresponding to an upstream motif or cis element capable of binding PEAl as set forth in Figure 1 at residues 3285-3298, a sequence corresponding to an upstream motif or cis element capable of binding ICS as set forth in Figure 1 at residues 3688-3699, a sequence corresponding to an upstream motif or cis element capable of binding ISGF2 as set forth in Figure 1 at residues 4170-4179, a sequence corresponding to an upstream motif or cis element capable of binding zinc as set forth in Figure 1 at residues 42&5-4293, a sequence corresponding to an upstream motif or cis element characteristic -of CAP/C:RP-galO as set forth in Figure *1 at residues 4379-4404, sequences corresponding to an upstream motif or cis element capable of binding APi as set forth in Figure 1 at residues 4428-4434, and 4627-4639, respectively, a sequence corresponding to an upstream motif or cis element capable of binding SRY as set forth in Figure 1 at residues 4625-4634, a sequence corresponding to an upstream motif or cis element characteristic of GC2 as set forth in Figure 1 at residues 4678-4711, a sequence corresponding to an upstream motif or cis element capable of binding PEA3 as set forth in Figure 1 at residues 4765-4769, a sequence corresponding to an upstream motif or cis element capable of MIR as set forth in Figure 1 at residues 4759-4954, a sequence corresponding to an upstream motif or cis element capable of binding NF-HNF-1 as set forth in Figure 1 at residues 4923-4941, a sequence corresponding to a thyroid receptor upstream motif or cis element as set forth in Figure 1 at residues 5151-5156, and a sequence corresponding to an upstream motif or cis element capable of binding N~hcB as set forth in Figure 1 at residues 5166-5175. For such purpose, the oligonucleotides must be capable of specifically hybridizing to a nucleic add molecule genetically or physically linked to the TIGR gene. As used herein, the term 'linked" refers to genetically, physically or operably linked.

As used herein, two nucleic add molecules are said to be capable of specifically hybridizing to one another if the two molecules are capable of forming an anti-parallel, double-stranded nudleic add structure, whereas they are unable to form a double-stranded structure when incubated with a non-TIGR nucleic acd *.20 molecule. A nucleic add molecule is said to be the "complement" of another nucleic adid molecule if they exhibit complete coniplementarity. As used herein, molecules '.:are said to exhibit "complete complementarity" when every nucleotide of one of the molecules is complementary to a nucleotide of the other. Two molecules are said to be "minimally complementary" if they can hybridize to one another with sufficient 25 stability to permit them to remain annealed to one another under at least conventional "low-sringency" conditions. Similarly, the molecules are said to be "complementary" if they can hybridize to one another with sufficient stability to permit them to remain annealed to one another under conventional "highstringency" conditions. Conventional stringency conditions are described by Sambrook, et al, (In. Molecular Cloning, a Laboratory Manual, 2nd Edition, Cold Spring Harbor Press, Cold Spring Harbor, New York (1989)), and by Haymes, et al (1n- Nuclec Adid Hybridizato, A Practical Approah, MEL Press, Washington, DC (1985)), both herein incorporated by reference). Departures from complete complementarity are therefore permissible, as long as such departures do not 35 completely preclude the capacity of the molecules to form a double-stranded structure. Thus, in order for an oligonucleotide to serve as a primer it need only be sufficiently complementary in sequence to be able to form a stable double-stranded structure under the particular solvent and salt concentrations employed.

Apart from their diagnostic or prognostic uses, such oligonucleotides may be employed to obtain other TIGR nucleic acid molecules. Such molecules include the TIGR-encoding nucleic acid molecule of non-human animals (particularly, cats, monkeys, rodents and dogs), fragments thereof, as well as their promoters and flanking sequences. Such molecules can be readily obtained by using the abovedescribed primers to screen cDNA or genomic libraries obtained from non-human species. Methods for forming such libraries are well known in the art. Such analogs may differ in their nucleotide sequences from that of SEQ ID NO. 1, SEQ ID NW. 2, SEQ ID NO: 3, SEQ ID NO. 4, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO: 7, SEQ ID NO. 8,SEQ ID NO. 9,SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO. 14, SEQ ID NO, 15, SEQ ID NO. 16, SEQ ID NO: 17, SEQ ID NO. 18, SEQ ID NO- 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO. 23, SEQ ID NO: 24, SEQ ID NO: 25, or from molecules consisting of sequences corresponding to an upstream motif or cis element characteristic of PRL-FP111 as set forth in Figure 1 at residues 370-388, and 4491-4502, respectively, a sequence corresponding to an upstream motif or cis element capable of binding GR/PR as set forth in Figure I at residues 433-445, sequences corresponding to an upstream shear stress motif or cis element as set forth in Figure 1 at residues 446-451, 1288-1293, 3597-3602, 4771-4776, and 5240-5245, respectively, sequences corresponding to glucocorticoid response upstream motif or cis element as set forth in Figure 1 at residues 574-600, 1042-1056, 2444-2468, 2442-2269, 3536-3563, 4574-4593, 4595-4614, 4851-4865,4844-4864,5079-5084, 5083-5111, respectively, a sequence corresponding to an upstream motif or cis element capable of binding CBE as set forth in Figure 1 at residues 735-746, a sequence corresponding to an upstream motif or cis element capable of binding NFE as set forth in Figure 1 at residues 774-795, a sequence corresponding to an upstream motif or cis element capable of binding KTF.1-CS as set forth in Figure 1 at residues 843-854, a sequence corresponding to an upstream motif or cis element capable of binding PRE is set forth in Figure 1 at residues 987- 1026, a sequence corresponding to an upstream motif or cis element capable of binding ETF-EGFR as set forth in Figure 1 at residues 1373-1388, a sequence corresponding to an upstream motif or cis element capable of binding SRE-cFos as set forth in Figure 1 at residues 1447-1456, a sequence corresponding to an upstream motif or cis element capable of binding Alu. as set forth in Figure 1 at residues 1331- 1550, a sequence corresponding to an upstream motif or cis element capable of binding VBP as set forth in Figure 1 at residues 1786-1797, a sequence corresponding to an upstream motif or cis element capable of binding Malt-CS as set forth in Figure 1 at residues 1832-1841, sequences corresponding to an upstream motif or cis element capable of binding ERE as set forth in Figure 1 at residues 2167-2195,3413- 3429, and 3892-3896, respectively, a sequence corresponding to an upstream motif or cis element capable of binding NF-mutagen as set forth in Figure 1 at residues 2329- 2338, a sequence corresponding to an upstream motif or cis element capable of binding myc-PRF as set forth in Figure 1 at residues 2403-2416, sequences corresponding to an upstream motif or cis element capable of binding A1'2 as set forth in Figure 1 at residues 2520-2535 and 5170-5187, respectively, sequences corresponding to an upstream motif or cis element capable of binding US IF as set forth in Figure 1 at residues 2622-2635, and 5105-5132, respectively, a sequence corresponding to an upstream motif or cis element characteristic of SBF as set forth in Figure 1 at residues 2733-2743, sequences corresponding to an upstream motif or cis element capable of binding NF-1 as set forth in Figure 1 at residues 2923-2938, 4144-4157, and 4887-4900, respectively, a sequence corresponding to an upstream motif or cis element capable of binding NF-MHCIIA/B as set forth in Figure 1 at 20 residues 2936-2944, a sequence corresponding to an upstream motif or cis element capable of binding PEAl as set forth in Figure 1 at residues 3285-3298, a sequence :1;':corresponding to an upstream motif or cis element capable of binding ICS as set forth in Figure 1 at residues 3688-3699, a sequence corresponding to an upstream motif or cis element capable of binding ISGF2 as set forth in Figure 1 at residues 0% 25 4170-4179, a sequence corresponding to an upstream motif or cis element capable of binding zinc as set forth in Figure 1 at residues 4285-423, a sequence corresponding to an upstream motif or cis element characteristic of CAP/CERP-galO as set forth in Figure 1 at residues 4379-4404, sequences corresponding to an upstream motif or cis element capable of binding AP1 as set forth in Figure 1 at residues 4428-4434, and 4627-4639, respectively, a sequence corresponding to an upstream motif or cis element capable of binding SRY as set forth in Figure 1 at residues 4625-4634, a sequence corresponding to an upstream motif or cis element chAracteristic of GC2 as set forth in Figure 1 at residues 4678-4711, a sequence corresponding to an upstream motif or cis element capable of binding PEA3 as set forth in Figure 1 at residues 4765-4769, a sequence corresponding to an upstream motif or cis element capable of MIR as set forth in Figure 1 at residues 4759-4954, a sequence corresponding to an upstream motif or cis element capable of binding NF-HNF-1 as set forth in Figure 1 at residues 4923-4941, a sequence corresponding to a thyroid receptor upstream motif or cis element as set forth in Figure 1 at residues 5151-5156, and a sequence corresponding to an upstream motif or cis element capable of binding NFhB as set forth in Figure 1 at residues 5166-5175 because complete complementarity is not needed for stable hybridization. The TIGR nucleic acid molecules of the present invention therefore also include molecules that, although capable of specifically hybridizing with TIGR nucleic acid molecules may lack "complete complementarity." Any of a variety of methods may be used to obtain the above-described nucleic acid molecules (Elles, Methods in Molecular Medicine: Molecular Diagnosis of Genetic Diseases, Humana Press (1996), herein incorporated by reference). SEQ ID NO: 1, SEQ ID NO. 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO. 6, SEQ ID NO: 7, SEQ ID NO. 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID N. 11, SEQ ID NO: 12, SEQ ID N. 13, SEQ ID NO: 14, SEQ ID NO:. 15, SEQ ID NC. 16, SEQ ED NO: 17, SEQ ID NO. 18, SEQ ID NO: 19, SEQ ID NO. 20, SEQ ID NO: 21, SEQ ID NO. 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NC. 25, sequences corresponding to an upstream motif or cis element characteristic of PRL-FP111 as set forth in Figure 1 e 20 at residues 370-388, and 4491-4502, respectively, a sequence corresponding to an e upstream motif or cis element capable of binding GR/PR as set forth in Figure I at residues 433-445, sequences corresponding to an upstream shear stress motif or cis element as set forth in Figure 1 at residues 446-451, 1288-1293, 3597-3602, 4771-4776, and 5240-5245, respectively, sequences corresponding to glucocorticoid response upstream motif or cis element as set forth in Figure 1 at residues 574-600,1042-1056, 2444-2468, 2442-2269, 3536-3563, 4574-4593, 4595-4614, 4851-4865, 4844-4864, 5079- 5084,5083-5111, respectively, a sequence corresponding to an upstream motif or cis element capable of binding CBE as set forth in Figure 1 at residues 735-746, a sequence corresponding to an upstream motif or cis element capable of binding NFE as set forth in Figure 1 at residues 774-795, a sequence corresponding to an upstream motif or cis element capable of binding KTF.1-CS as set forth in Figure 1 at residues 843-854, a sequence corresponding to an upstream motif or cis element capable of binding PRE is set forth in Figure 1 at residues 987-1026, a sequence corresponding to an upstream motif or cis element capable of binding ET-EGFR as set forth in Figure I at residues 1373-1388, a sequence corresponding to an upstream motif or cis element capable of binding SRE-cFos as set forth in Figure 1 at residues 1447-1456, a sequence corresponding to an upstream -motif or cis element capable of binding Alu as set forth in Figure 1 at residues 1331-1550, a sequence corresponding to an upstream motif or cis element capable of binding VBP as set forth in Figure 1 at residues 1786-1797, a sequence corresponding to an upstream motif or cis element capable of binding Malt-CS as set forth in Figure 1 at residues 1832-1841, sequences corresponding to an upstream motif or cis element capable of binding ERE as set forth in Figure 1 at residues 2167-2195, 3413-3429, and 3892-3896, respectively, a sequence corresponding to an upstream motif or cis element capable of binding NF-mutagen as set forth in Figure 1 at residues 2329-2338, a sequence corresponding to an upstream motif or cis element capable of binding myc-PEF as set forth in Figure 1 at residues 2403-2416, sequences corresponding to an upstream motif or cis element capable of binding AP2 as set forth in Figure 1 at residues 2520- 2535 and 5170-5187, respectively, sequences corresponding to an upstream motif or cis element capable of binding HSTF as set forth in Figure 1 at residues 2622-2635, and 5105-5132, respectively, a sequence corresponding to an upstream motif or cis element characteristic of SUF as set forth in Figure 1 at residues 2733-2743, sequences corresponding to an upstream motif or cis element capable of binding NF-1 as set forth in Figure 1 at residues 2923-2938, 4144-4157, and 4887-4900, respectively, a 20 sequence corresponding to an upstream motif or cis element capable of binding NF- MHCIIA/B as set forth in Figure 1 at residues 2936-2944, a sequence corresponding to an upstream motif or cis element capable of binding PEAl as set forth in Figure 1 at residues 3285-3298, a sequence corresponding to an upstream motif or cis element capable of binding ICS as set forth in Figure 1 at residues 3688-3699, a sequence corresponding to an upstream motif or cis element capable of binding NAMF as set forth in Figure 1 at residues 4170-4179, a sequence corresponding to an upstream motif or cis element capable of binding zinc as set forth in Figure 1 at residues 42&5-4293, a sequence corresponding to an upstream motif or cis element characteristic of CAP/CEP-galO as set forth in Figure 1 at residues 4379-4404, sequences corresponding to an upstream motif or cis element capable of binding APi as set forth in Figure 1 at residues 4428-4434, and 4627-4639, respectively, a sequence corresponding to an upstream motif or cis element capable of binding SRY as set forth in Figure 1 at residues 4625-4634, a sequence corresponding to an upstream motif or cis element characteristic of GC2 as set forth in Figure 1 at residues 4678-4711, a sequence corresponding to an upstream motif or cis element capable of binding PEA3 as set forth in Figure 1 at residues 4765-4769, a sequence corresponding to an upstream motif or cis element capable of MIR as set forth in Figure 1 at residues 4759-4954, a sequence corresponding to an upstream motif or cis element capable of binding NF-HNF-1 as set forth in Figure 1 at residues 4923-4941, a sequence corresponding to a thyroid receptor upstream motif or cis element as set forth in Figure 1 at residues 5151-5156, and a sequence corresponding to an upstream motif or cis element capable of binding NFKB as set forth in Figure 1 at residues 5166-5175 may be used to synthesize all or any portion of the TIGR promoter or any of the TIGR upstream motifs or portions the TIGR cDNA (Zamechik et al., Proc. Natl. Acad. Sci. 83:4143 (1986); Goodchild et Proc.

Natl. Acad. Sci. 85:5507 (1988); Wickstrom et al., Proc. Natl. Acad. Sci. 85:1028; Holt, J.T. et al., Molec. Cell Biol. 8:963 (1988); Gerwirtz, AM. et al., Science 242:1303 (1988); Anfossi, et al., Proc. Natl. Acad. Sci. 86:3379 (1989); Becker, et al., EMBO 1. 8:3679 (1989); all of which references are incorporated herein by reference).

S:Automated nucleic acid synthesizers may be employed for this purpose. In lieu of such synthesis, the disclosed SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO:. 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO. 8, SEQ ID NO: S 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:. 14, 20 SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO:. 17, SEQ ID NO: 18, SEQ ID NO:. 19, SEQ ID NO:. 20, SEQ ID NO: 21, SEQ ID NO. 22, SEQ ID NO: 23, SEQ ID NO:. 24, SEQ ID NO: 25, sequences corresponding to an upstream motif or cis element e" ['characteristic of PRL-FP111 as set forth in Figure 1 at residues 370-388, and 4491- 4502, respectively, a sequence corresponding to an upstream motif or cis element capable of binding GR/PR as set forth in Figure 1 at residues 433-445, sequences corresponding to an upstream shear stress motif or cis element as set forth in Figure 1 at residues 446-451,1288-1293,3597-3602,4771-4776, and 5240-5245, respectively, sequences corresponding to glucocorticoid response upstream motif or cis element as set forth in Figure 1 at residues 574-600, 1042-1056, 2444-2468, 2442-2269, 3536- 3563,4574-4593,4595-4614,4851-4865,4844-4864,5079-5084, 5083-5111, respectively, a sequence corresponding to an upstream motif or cis element capable of binding CBE as set forth in Figure 1 at residues 735-746, a sequence corresponding to an upstream motif or cis element capable of binding NFE as set forth in Figure 1 at residues 774-795, a sequence corresponding to an upstream motif or cis element capable of binding KTF.1-CS as set forth in Figure 1 at residues 843-854, a sequence corresponding to an upstream motif or cis element capable of binding PRE is set forth in Figure 1 at residues 987-1026, a sequence corresponding to an upstream motif or cis element capable of binding ETF-EGFR as set forth in Figure 1 at residues 1373-1388, a sequence corresponding to an upstream motif or cis element capable of binding SRE-cFos as set forth in Figure 1 at residues 1447-1456, a sequence corresponding to an upstream motif or cis element capable of binding Alu as set forth in Figure 1 at residues 1331-1550, a sequence corresponding to an upstream motif or cis element capable of binding VBP as set forth in Figure 1 at residues 1786-1797, a sequence corresponding to an upstream motif or cis element capable of binding Malt-CS as set forth in Figure 1 at residues 1832-1841, sequences corresponding to an upstream motif or cis element capable of binding ERE as set forth in Figure 1 at residues 2167-2195, 3413-3429, and 3892-3896, respectively, a sequence corresponding to an upstream motif or cis element capable of binding NFmutagen as set forth in Figure 1 at residues 2329-2338, a sequence corresponding to an upstream motif or cis element capable of binding myc-PRF as set forth in Figure 1 at residues 2403-2416, sequences corresponding to an upstream motif or cis element capable of binding AP2 as set forth in Figure 1 at residues 2520-2535 and 5170-5187, respectively, sequences corresponding to an upstream motif or cis element capable of binding HST as set forth in Figure 1 at residues 2622-2635, and o 20 .5105-5132, respectively, a sequence corresponding to an upstream motif or cis element characteristic of SBF as set forth in Figure 1 at residues 2733-2743, sequences ool. corresponding to an upstream motif or cis element capable of binding NP-i as set forth in Figure 1 at residues 2923-2938, 4144-4157, and 4887-4900, respectively, a sequence corresponding to an upstream motif or cis element capable of binding NP- MHCII.A/B as set forth in Figure 1 at residues 2936-2944, a sequence corresponding 00 to an upstream motif or cis element capable of binding PEAl as set forth in Figure 1 at residues 3285-3298, a sequence corresponding to an upstream motif or cis element capable of binding ICS as set forth in Figure 1 at residues 3688-3699, a sequence corresponding to an upstream motif or cis element capable of binding ISGF as set forth in Figure 1 at residues 4170-4179, a sequence corresponding to an upstream motif or cis element capable of binding zinc as set forth in Figure 1 at residues 4285-4293, a sequence corresponding to an upstream motif or cis element characteristic of CAP/CRP-galO as set forth in Figure 1 at residues 4379-4404, sequences corresponding to an upstream motif or cis element capable of binding AP1 as set forth in Figure 1 at residues 4428-4434, and 4627-4639, respectively, a sequence corresponding to an upstream motif or cis element capable of binding SRY as set forth in Figure 1 at residues 4625-4634, a sequence corresponding to an upstream motif or cis element characteristic of GC2 as set forth in Figure 1 at residues 4678-4711, a sequence corresponding to an upstream motif or cis element capable of binding PEA3 as set forth in Figure I at residues 4765-4769, a sequence corresponding to an upstream motif or cis element capable of MIR as set forth in Figure 1 at residues 4759-4954, a sequence corresponding to an upstream motif or cis element capable of binding NF-HNF-1 as set forth in Figure 1 at residues 4923-4941, a sequence corresponding to a thyroid receptor upstream motif or cis element as set forth in Figure 1 at residues 5151-5156, and a sequence corresponding to an upstream motif or cis element capable of binding NFB as set forth in Figure 1 at residues 5166-5175 may be used to define a pair of primers that can be used with the polymerase chain reaction (Mullis, K. et al., Cold Spring Harbor Symp. Quant. Biol.

51:263-273 (1986); Erlich H. et al., EP 50,424; EP 84,796, EP 258,017, EP 237,362; Mullis, EP 201,184; Mullis K. et al., US 4,683,202; Erlich, FL, US 4,582,788; and Saiki, R. et al, US 4,683,194)) to amplify and obtain any desired TIGR gene DNA molecule or fragment The TIGR promoter sequence(s) and TIGR flanking sequences can also be obtained by incubating oligonudeotide probes of TIGR oligonucleotides with members of genomic human libraries and recovering clones that hybridize to the probes. In a second embodiment, methods of "chromosome walking," or 3' or RACE may be used (Frohman, M.A. eta., Proc. Natl. Acad. Sci. 85:8998-9002 (1988), herein incorporated by reference); Ohara, 0. et al., Proc. Natl. Acad. Sci.

86:5673-5677 (1989), herein incorporated by reference) to obtain such sequences.

IL Uses of the Molecules of the Invention in the Diagnosis and Prognosis of Glaucoma and Related Diseases A particularly desired use of the present invention relates to the diagnosis of glaucoma, POAG, pigmentary glaucoma, high tension glaucoma and low tension glaucoma and their related diseases. Another particularly desired use of the present invention relates to the prognosis of glaucoma, POAG, pigmentary glaucoma, high tension glaucoma and low tension glaucoma and their related diseases. As used herein the term "glaucoma" includes both primary glaucomas, secondary glaucomas, juvenile glaucomas, congenital glaucomas, and familial glaucomas, including, without limitation, pigmentary glaucoma, high tension glaucoma and low tension glaucoma and their related diseases. As indicated above, methods for diagnosing or prognosing glaucoma suffer from inaccuracy, or require multiple examinations. The molecules of the present invention may be used to define superior assays for glaucoma. Quite apart from such usage, the molecules of the present invention may be used to diagnosis or predict an individual's sensitivity to elevated intraocular pressure upon administration of steroids such as glucocorticoids or corticosteroids, or anti-inflammatory steroids). Dexamethasone, cortisol and prednisolone are preferred steroids for this purpose. Medical conditions such as inflammatory and allergic disorders, as well as organ transplantation recipients, benefit from treatment with glucocorticoids. Certain individuals exhibit an increased sensitivity to such steroids "steroid sensitivity"), which is manifested by an undesired increase in intraocular pressure.

The present invention may be employed to diagnosis or predict such sensitivity, as well as glaucoma and related diseases.

In a first embodiment, the TIGR molecules of the present invention are used to determine whether an individual has a mutation affecting the level the concentration of TIGR mRNA or protein in a sample, etc.) or pattern the kinetics of expression, rate of decomposition, stability profile. etc.) of the 11GR 20 expression (collectively, the '71GR response" of a cell or bodily fluid) (for example, a mutation in the TIGR gene, or in a regulatory region(s) or other gene(s) that control or affect the expression of TIGR), and being predictive of individuals who would be predisposed to glaucoma (prognosis), related diseases, or steroid sensitivity. As used herein, the 11CR response manifested by a cell or bodily fluid is saidto be"altered" if it differs from the 11CR response of cells or of bodily fluids of normal individuals. Such alteration may be manifested by either abnormally increased or abnormally diminished 11CR response. To determine whether a 11CR response is altered, the 11CR response manifested by the cell or bodily fluid of the patient is compared with that of a similar cell sample (or bodily fluid sample) of normal individuals. As will be appreciated, it is not necessary to re-determine the 11CR response of the cell sample (or bodily fluid sample) of normal individuals each time such a comparison is made; rather, the 11CR response of a particular individual may be compared with previously obtained values of normal individuals.

In one sub-embodiment, such an analysis is conducted by determining the presence and/or identity of polymorphism(s) in the TIGR gene or its flanking regions which are associated with glaucoma, or a predisposition (prognosis) to glaucoma, related diseases, or steroid sensitivity. As used herein, the term "TMGR flanking regions" refers to those regions which are located either upstream or downstream of the TIGR coding region.

Any of a variety of molecules can be used to identify such polymorphiism(s).

in one embodiment, SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO, 4, SEQ ID) SEQ ID NO: 6, SEQD N,7,SEQ DN,8, SEQ DN.9, Q SEQ ID Na. 11, SEQ ID NO: 12, SEQ ID NO, 13, SEQ ID NO: 14, SEQ ID NO, SEQ ID NO, 16, SEQ ID NO: 17, SEQ ID NO 18, SEQ ID NO: 19, SEQIED SEQ MD NO: 21, SEQ ID NO: 22, SEQ ID 23, SEQ ID NO: 24, SEQ ID NO: sequences corresponding to an upstream motif or cis element characteristic of PEL- FF111 as set forth in Figure 1 at residues 370-388, and 4491-4502, respectively, a sequence corresponding to an upstream motif or cis element capable of binding GRIPR as set forth in Figure 1 at residues 433-445, sequences corresponding to an upstream shear stress motif or cis element as set forth in Figure 1 at residues 446- 451, 1288-1293, 3597-3602, 4771-4776, and 5240-5245, respectively, sequences corresponding to glucocorticoid response upstream motif or cis element as set forth in Figure 1 at residues 574-600, 1042-1056, 2444-2468, 2442-2269, 3536-3563, 4574- 4593,4595-4614,4851-4865,4844-4864,5079-5084, 5083-5111, respectively, a sequence 20 corresponding to an upstream motif or cis element capable of binding CBE as set forth in Figure 1 at residues 735-746, a sequence corresponding to an upstream motif ::or cis element capable of binding NFE as set forth in Figure 1 at residues 774-795, a sequence corresponding to an upstream motif or cis element capable of binding KTF.1-CS as set forth in Figure 1 at residues 843-854, a sequence corresponding to an upstream motif or cis element capable of binding PRE is set forth in Figure 1 at residues 987-1026, a sequence corresponding to an upstream motif or cis element capable of binding ETF-EGFR as set forth in Figure 1 at residues 1373-1388, a .sequence corresponding to an upstream motif or cis ele *ment capable of binding SRE-c~os as set forth in Figure 1 at residues 1447-1456, a sequence corresponding to an upstream motif or cis element capable of binding Alu as set forth in Figure 1 at residues 1331-1550, a sequence corresponding to an upstream motif or cis element capable of binding VBP as set forth in Figure 1 at residues 1786-1797, a sequence corresponding to an upstream motif or cis element capable of binding Malt-CS as set forth in Figure 1 at residues 1832-1841, sequences corresponding to an upstream motif or cis element capable of binding EKE as set forth in Figure 1 at residues 2167- 2195, 3413-3429, and 3892-3896, respectively, a sequence corresponding to an upstream motif or cis element capable of binding NF-mutagen as set forth in Figure 1 at residues 2329-2338, a sequence corresponding to an upstream motif or cis element capable of binding myc-PRF as set forth in Figure 1 at residues 2403-2416, sequences corresponding to an upstream motif or cis element capable of binding AP2 as set forth in Figure 1 at residues 2520-2535 and 5170-5187, respectively, sequences corresponding to an upstream motif or cis element capable of binding HSTF as set forth in Figure 1 at residues 2622-2635, and 5105-5132, respectively, a sequence corresponding to an upstream motif or cis element characteristic of SBF as set forth in Figure 1 at residues 2733-2743, sequences corresponding to an upstream motif or cis element capable of binding NF-1 as set forth in Figure 1 at residues 2923-2938, 4144-4157, and 4887-4900, respectively, a sequence corresponding to an upstream motif or cis element capable of binding NF-MHCIIA/B as set forth in Figure 1 at residues 2936-2944, a sequence corresponding to an upstream motif or cis element capable of binding PEAl as set forth in Figure 1 at residues 3285-3298, a sequence corresponding to an upstream motif or cis element capable of binding ICS as set forth in Figure 1 at residues 3688-3699, a sequence corresponding to an upstream motif or cis element capable of binding ISMF as set forth in Figure 1 at residues 4170-4179, a sequence corresponding to an upstream motif or cis element 20 capable of binding zinc as set forth in Figure 1 at residues 4285-4293, a sequence corresponding to an upstream motif or cis element characteristic of CAP, CEP-gaIO as set forth in Figure 1 at residues 4379-4404, sequences corresponding to an upstream motif or cis element capable of binding AP1 as set forth in Figure 1 at residues 4428-4434, and 4627-4639, respectively, a sequence corresponding to an upstream motif or cis element capable of binding SRY as set forth in Figure 1 at residues 4625-4634, a sequence corresponding to an upstream motif or cis element characteristic of GC2 as set forth in Figure 1 at residues 4678-4711, a sequence corresponding to an upstream motif or cis element capable of binding PEA3 as set forth in Figure 1 at residues 4765-4769, a sequence corresponding to an upstream motif or cis element capable of MIR as set forth in Figure 1 at residues 4759-4954, a sequence corresponding to an upstream motif or cis element capable of binding NF- HNF-1 as set forth in Figure 1 at residues 4923-4941, a sequence corresponding to a thyroid receptor upstream motif or cis element as set forth in Figure 1 at residues 5151-5156, and a sequence corresponding to an upstream motif or cis element capable of binding NFOc as set forth in Figure 1 at residues 5166-5175 (or a subsequence thereof) may be employed as a marker nucleic acid molecule to identify such polymorphism(s).

Alternatively, such polymorphisms can be detected through the use of a marker nucleic acid molecule or a marker protein that is genetically linked to a polynucleotide that co-segregates with) such polymorphism(s). As stated above, the TIGR gene and/or a sequence or sequences that specifically hybridize to the TIGR gene have been mapped to chromosome lq, 21-32, and more preferably to the TIGR gene located at chromosome 1, q21-27, and more preferably to the TIGR gene located at chromosome 1, q22-26, and most preferably to the TIGR gene located at chromosome 1, q24. In a preferred aspect of this embodiment, such marker nucleic acid molecules will have the nudeotide sequence of a polynucleotide that is closely genetically linked to such polymorphism(s) markers located at chromosome 1, q19-25 (and more preferably chromosome 1, q23-25, and most preferably chromosome 1, q24.

Localization studies using a Stanford G3 radiation hybrid panel mapped the TIGR gene with the D1S2536 marker nucleic acid molecules at the D1S2536 locus with a LOD score of 6.0. Other marker nucleic acid molecules in this region include: D1S210; D1S1552; D1S2536; D1S2790; SHGC-12820; and D1S2558. Other polynucleotide markers that map to such locations are known and can be employed to identify such polymorphism(s).

The genomes of animals and plants naturally undergo spontaneous mutation in the course of their continuing evolution (Gusella, Ann. Rev. Biochem. 55:831- 854 (1986)). A "polymorphism" in the TIGR gene or its flanking regions is a variation or difference in the sequence of the TIGR gene or its flanking regions that arises in some of the members of a species. The variant sequence and the "original" sequence co-exist in the species' population. In some instances, such co-existence is in stable or quasi-stable equilibrium.

.A polymorphism is thus said to be "allelic," in that, due to the existence of the polymorphism, some members of a species may have the original sequence (Le.

the original "allele") whereas other members may have the variant sequence the variant "allele"). In the simplest case, only one variant sequence may exist, and the polymorphism is thus said to be di-allelic. In other cases, the species' population may contain multiple alleles, and the polymorphism is termed tri-allelic, etc. A single gene may have multiple different unrelated polymorphisms. For example, it may have a di-allelic polymorphism at one site, and a multi-allelic polymorphism at another site.

The variation that defines the polymorphism may range from a single nucleotide variation to the insertion or deletion of extended regions within a gene.

In some cases, the DNA sequence variations are in regions of the genome that are characterized by short tandem repeats (STRs) that include tandem di- or trinucleotide repeated motifs of nuceotides. Polymorphisms characterized by such tandem repeats are referred to as "variable number tandem repeat" ("VNTR") polymorphisms. VNTRs have been used in identity and paternity analysis (Weber, J.L, U.S. Patent 5,075,217; Armour, J.A.L et al., FEBS Lett. 307:113-115 (1992); Jones, L. et al., Eur. I. Haematol. 39:144-147 (1987); Horn, G.T. et al., PCT Application WO91/14003; Jeffreys, European Patent Application 370,719; Jeffreys, U.S.

Patent 5,175,082); Jeffreys. A.J. et al., Amer. J. Hum. Genet. 39:11-24 (1986); Jeffreys.

AJ. et al., Nature 316:76-79 (1985); Gray, LC. et al., Proc. R. Acad. Soc. Lond. 243:241- 253 (1991); Moore, S.S. et al., Genomics 10:654-660 (1991); Jeffreys, A.J. et al.,Anim.

S.:Genet. 18:1-15 (1987); Hillel, J. et al, Anim. Genet. 20:145-155 (1989); Hillel, J. et al., Genet. 124:783-789 (1990)).

In an alternative embodiment, such polymorphisms can be detected through the use of a marker nucleic acid molecule that is physically linked to such 20 polymorphism(s). For this purpose, marker nucleic acid molecules comprising a nucleotide sequence of a polynucleotide located within 1 mb of the polymorphism(s), and more preferably within 100 kb of the polymorphism(s), and most preferably within 10 kb of the polymorphism(s) can be employed. Examples of such marker nucleic acids are set out in SEQ ID NO. 1, SEQ ID NO 2, SEQ ID N.O 3, SEQ ID NO 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID o NO SEQ ID N 10, SEQ ID N. 11, SEQ ID NO: 12, SEQ ID NO: 13,SEQ IDNO: 14, SEQ ID N.O 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: In another embodiment a marker nucleic acid will be used that is capable of specifically detecting TIGRmtl, TIGRmt2, TIGRmt3, TIGRmt4, TIGRmt5, TIGRsvl,or a combination of these mutations. Methods to detect base(s) substitutions, base(s) deletions and base(s) additions are known in the art methods to genotype an individual). For example, "Genetic Bit Analysis method is disclosed by Goelet, P. et al., WO 92/15712, herein incorporated by reference, may be used for detecting the single nucleotide polymorphisms of the present invention. GBA is a method of polymorphic site interrogation in which -the nucleotide sequence information surrounding the site of variation in a target DNA sequence is used to design an oligonuceotide primer that is complementary to the region immediately adjacent to, but not including, the variable nucleotide in the target DNA. The target DNA template is selected from the biological sample and hybridized to the interrogating primer. This primer is extended by a single labeled dideoxynucleotide using DNA polymerase in the presence of two, and preferably all four chain terminating nucleoside triphosphate precursors. Cohen, D. et al., (PCT Application W091/02087) describes a related method of genotyping.

Other primer-guided nucleotide incorporation procedures for assaying polymorphic sites in DNA have been described (Komher, J. S. et al., Nucl. Acids. Res.

17:7779-7784 (1989), herein incorporated by reference; Sokolov, B. Nucl. Acids Res.

18:3671 (1990), herein incorporated by reference; Syviinen, et al., Genomics 8:684 692 (1990), herein incorporated by reference; Kuppuswamy, M.N. et al., Proc.

Natl. Acad. Sci 88:1143-1147 (1991), herein incorporated by reference; Prezant, T.R et al., Hum. Mutat. 1:159-164 (1992), herein incorporated by reference; Ugozzoli, L et al., GATA 9:107-112 (1992), herein incorporated by reference; Nyrdn, P. et al., Anal. Biochem. 208:171-175 (1993), herein incorporated by reference).

The detection of polymorphic sites in a sample of DNA may be facilitated through the use of nucleic acid amplification methods. Such methods specifically increase the concentration of polynucleotides that span the polymorphic site, or include that site and sequences located either distal or proximal to it. Such amplified molecules can be readily detected by gel electrophoresis or other means.

Another preferred method of achieving such amplification employs the polymerase chain reaction (Mullis, K et aL, Cold Spring Harbor Symp. Quant.

Biol. 51:263-273 (1986); Erlich H. et al., European Patent Appln. 50,424; European Patent Appln. 84,796, European Patent Application 258,017, European Patent Appln.

237,362; Mullis, K, European Patent Appln. 201,184; Mullis K et al., US. Patent No.

4,683,202; Erlich, US. Patent No. 4,582,788; and Saiki, R. et al., US. Patent No.

4,683,194), using primer pairs that are capable of hybridizing to the proximal sequences that define a polymorphism in its double-stranded form.

In lieu of PCR, alternative methods, such as the "Ligase Chain Reaction" may be used (Barany, Proc. Natl. Acad. Sci. 88:189-193 (1991).

LCR uses two pairs of oligonucleotide probes to exponentially amplify a specific target. The sequences of each pair of oligonucleotides is selected to permit the pair to hybridize to abutting sequences of the same strand of the target. Such hybridization forms a substrate for a template-dependent ligase. As with PCR, the resulting products thus serve as a template in subsequent cycles and an exponential amplification of the desired sequence is obtained.

LCR can be performed with oligonucleotides having the proximal and distal sequences of the same strand of a polymorphic site. In one embodiment, either oligonucleotide will be designed to include the actual polymorphic site of the polymorphism. In such an embodiment, the reaction conditions are selected such that the oligonucleotides can be ligated together only if the target molecule either contains or lacks the specific nucleotide that is complementary to the polymorphic site present on the oligonucleotide. Alternatively, the oligonucleotides may be selected such that they do not include the polymorphic site (see, Segev, PCT Application WO 90/01069).

The "Oligonucleotide Ligation Assay" may alternatively be employed (Landegren, U. et al., Science 241:1077-1080 (1988)). The OLA protocol uses two oligonucleotides which are designed to be capable of hybridizing to abutting sequences of a single strand of a target. OLA, like LCR, is particularly suited for the detection of point mutations. Unlike LCR, however, OLA results in 20 "linear" rather than exponential amplification of the target sequence.

Nickerson, D.A. et al., have described a nucleic acid detection assay that combines attributes of PCR and OLA (Nickerson, D.A. et al., Proc. Natl. Acad. Sci.

87:8923-8927 (1990). In this method, PCR is used to achieve the exponential amplification of target DNA, which is then detected using OLA. In addition to S 25 requiring multiple, and separate, processing steps, one problem associated with such combinations is that they inherit all of the problems associated with PCR and

OLA.

Schemes based on ligation of two (or more) oligonucleotides in the presence of nucleic acid having the sequence of the resulting "di-oligonuceotide", thereby amplifying the di-oligonucleotide, are also known (Wu, D.Y. et al., Genomics 4:560 (1989)), and may be readily adapted to the purposes of the present invention.

Other known nucleic acid amplification procedures, such as allele-specific oligomers, branched DNA technology, transcription-based amplification systems, or isothermal amplification methods may also be used to amplify and analyze such polymorphisms (Malek, LT. et al., US. Patent 5,130,238; Davey, C et at, European Patent Application 329,822; Schuster et al., U.S. Patent 5,169,766; Miller, ILI. et al., PCT appln. WO 89/06700; Kwoh, D. et al., Proc. Natl. Acad. Sci. 86:1173 (1989); Gingeras, T.L et al., PCT application WO 88/10315; Walker, G.T. et al., Proc.

Natl. Acad. Sci. 89:392-396 (1992)). All the foregoing nucleic acid amplification methods could be used to predict or diagnose glaucoma.

The identification of a polymorphism in the TIGR gene can be determined in a variety of ways. By correlating the presence or absence of glaucoma in an individual with the presence or absence of a polymorphism in the TIGR gene or its flanking regions, it is possible to diagnose the predisposition (prognosis) of an asymptomatic patient to glaucoma, related diseases, or steroid sensitivity. If a polymorphism creates or destroys a restriction endonuclease cleavage site, or if it results in the loss or insertion of DNA a VNTR polymorphism), it will alter the size or profile of the DNA fragments that are generated by digestion with that restriction endonuclease. As such, individuals that possess a variant sequence can be distinguished from those having the original sequence by restriction fragment analysis. Polymorphisms that can be identified in this manner are termed restriction fragment length polymorphisms" FLPs"). RFLPs have been widely used in human and animal genetic analyses (Glassberg, UK patent Application 2135774; Skolnick, M.H et aL, Cytogen. Cell Genet. 32:58-67 (1982); Botstein, D. et al., Ann. J. Hum. Genet. 32:314-331 (1980); Fischer, S.G et al. (PCT Application W090/13668); Uhlen, PCT Application W090/11369)). The role of TIGR in glaucoma pathogenesis indicates that the presence of genetic alterations DNA polymorphisms) that affect the TIGR response can be employed to predict glaucoma A preferred method of achieving such identification employs the singlestrand conformational polymorphism (SSCP) approach. The SSCP technique is a method capable of identifying most sequence variations in a single strand of DNA, typically between 150 and 250 nucleotides in length (Elles, Methods in Molecular Medicine: Molecular Diagnosis of Genetic Diseases, Humana Press (1996), herein incorporated by reference); Onita et al., Genomics 5: 874-879 (1989), herein incorporated by reference). Under denaturing conditions a single strand of DNA will adopt a conformation that is uniquely dependent on its sequence conformation.

This conformation usually will be different, even if only a single base is changed.

Most conformations have been reported to alter the physical configuration or size sufficiently to be detectable by electrophoresis. A number of protocols have been described for SSCP including, but not limited to Lee et at., AnaL Biochem. 205:289-293 (1992), herein incorporated by reference; Suzuki et al., Anal. Biochem. 192: 82-84 (1991), herein incorporated by reference; Lo et al., Nucleic Acids Research 20: 1005- 1009 (1992), herein incorporated by reference; Sarkar et al., Genomics 13: 441-443 (1992), herein incorporated by reference).

In accordance with this embodiment of the invention, a sample DNA is obtained from a patient's cells. In a preferred embodiment, the DNA sample is obtained from the patient's blood. However, any source of DNA may be used. The DNA is subjected to restriction endonuclease digestion. TIGR is used as a probe in accordance with the above-described RFLP methods. By comparing the RFLP pattern of the TIGR gene obtained from normal and glaucomatous patients, one can determine a patient's predisposition (prognosis) to glaucoma. The polymorphism obtained in this approach can then be cloned to identify the mutation at the coding region which alters the protein's structure or regulatory region of the gene which affects its expression level. Changes involving promoter interactions with other regulatory proteins can be identified by, for example, gel shift assays using HTM cell extracts, fluid from the anterior chamber of the eye, serum, etc. Interactions of TIGR protein in glaucomatous cell extracts, fluid from the anterior chamber of the eye, serum, etc. can be compared to control samples to thereby identify changes in 20 those properties of TIGR that relate to the pathogenesis of glaucoma. Similarly such extracts and fluids as well as others (blood, etc.) can be used to diagnosis or predict .steroid sensitivity.

Several different classes of polymorphisms may be identified through such methods. Examples of such classes include: polymorphisms present in the TIGR cDNA of different individuals; polymorphisms in non-translated TIGR gene sequences, including the promoter or other regulatory regions of the TIGR gene; (3) polymorphisms in genes whose products interact with TIGR regulatory sequences; polymorphisms in gene sequences whose products interact with the TIGR protein, or to which the TIGR protein binds.

In an alternate sub-embodiment, the evaluation is conducted using oligonucleotide "probes" whose sequence is complementary to that of a portion of SEQ ID NO, 1, SEQ ID NO. 2 SEQ ID NO. 3, SEQ ID NO:. 4, or SEQ ID NO: 5. Such molecules are then incubated with cell extracts of a patient under conditions sufficient to permit nucleic acid hybridization.

In one sub-embodiment of this aspect of the present invention, one can diagnose or predict glaucoma, related diseases and steroid sensitivity by ascertaining the ICR response in a biopsy (or a macrophage or other blood cell sample), or other cell sample, or more preferably, in a sample of bodily fluid (especially, blood, serum, plasma, tears, buccal cavity, etc.). Since the T1GR gene is induced in response to the presence of glucocorticoids, a highly preferred embodiment of this method comprises ascertaining such TIGR response prior to, during and/or subsequent to, the administration of a glucocorticoid. Thus, by way of illustration, glaucoma could be diagnosed or predicted by determining whether the administration of a glucocorticoid (administered topically, intraocularly, intramuscularly, systemically, or otherwise) alters the TIGR response of a particular individual, relative to that of normal individuals. Most preferably, for this purpose, at least a "TIGR gene-inducing amount" of the glucocorticoid will be provided. As used herein, a 11GR gene-inducing amount of a glucocorticoid is an amount of glucocorticoid sufficient to cause a detectable induction of T1GR expression in cells of glaucomatous or non-glaucomatous individuals.

I. Methods of Administration The agents of the present invention can be formulated according to known methods to prepare pharmacologically acceptable compositions, whereby these materials, or their functional derivatives, having the desired degree of purity are combined in admixture with a physiologically acceptable carrier, excipient, or S"-stabilizer. Such materials are non-toxic to recipients at the dosages and *"concentrations employed. The active component of such compositions may be agents analogs or mimetics of such molecules. Where nucleic acid molecules are employed, such molecules may be sense, antisense or triplex oligonucleotides of the 11GR promoter, 11GR cDNA, T1GR intron, 11GR exon or T1GR gene.

A composition is said to be "pharmacologically acceptable" if its administration can be tolerated by a recipient patient. An agent is physiologically significant if its presence results in a detectable change in the physiology of a recipient patient Suitable vehicles and their formulation, inclusive of other human proteins, human serum albumin, are described, for example, in Remington's Pharmaceutical Sciences (16th ed., Osol, Ed., Mack, Easton PA (1980)).

If the composition is to be water soluble, it may be formulated in a buffer such as phosphate or other organic acid salt preferably at a pH of about 7 to 8. If the composition is only partially soluble in water, it may be prepared as a microemulsion by formulating it with a nonionic surfactant such as Tween, Pluronics, or PEG, Tween 80, in an amount of, for example, 0.04-0.05% to increase its solubility. The term "water soluble" as applied to the polysaccharides and polyethylene glycols is meant to include colloidal solutions and dispersions. In general, the solubility of the cellulose derivatives is determined by the degree of substitution of ether groups, and the stabilizing derivatives useful herein should have a sufficient quantity of such ether groups per anhydroglucose unit in the cellulose chain to render the derivatives water soluble. A degree of ether substitution of at least 035 ether groups per anhydroglucose unit is generally sufficient Additionally, the cellulose derivatives may be in the form of alkali metal salts, for example, the Li, Na, K or Cs salts.

Optionally other ingredients may be added such as antioxidants, e.g., ascorbic acid; low molecular weight (less than about ten residues) polypeptides, e.g., polyarginine or tripeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinyl pyrrolidone; amino acids, such as glycine, glutamic add, aspartic acid, or arginine; monosaccharides, 20 disaccharides, and other carbohydrates including cellulose or its derivatives, glucose, mannose, or dextrins; chelating agents such as EDTA; and sugar alcohols such as mannitol or sorbitoL Additional pharmaceutical methods may be employed to control the duration of action. Controlled or sustained release preparations may be achieved through the use of polymers to complex or absorb the TIGR molecule(s) of the composition. The controlled delivery may be exercised by selecting appropriate macromolecules (for example polyesters, polyamino acids, polyvinyl pyrrolidone, ethylenevinylacetate, methylcellulose, carboxymethylcellulose, or protamine sulfate) and the concentration of macromolecules as well as the methods of incorporation in order to control release.

Sustained release formulations may also be prepared, and include the formation of microcapsular particles and implantable articles. For preparing sustained-release compositions, the TIGR molecule(s) of the composition is preferably incorporated into a biodegradable matrix or microcapsule. A suitable material for this purpose is a polylactide, although other polymers of poly-(ahydroxycarboxylic adds), such as poly-D-(-)-3-hydroxybutyric acid (EP 133,988A), can be used. Other biodegradable polymers include poly(lactones), poly(orthoesters), polyamino acids, hydrogels, or poly(orthocarbonates) poly(acetals). The polymeric material may also comprise polyesters, poly(lactic acid) or ethylene vinylacetate copolymers. For examples of sustained release compositions, see U.S. Patent No. 3,773,919, EP 58,481A, U.S. Patent No. 3,887,699, EP 158,277A, Canadian Patent No. 1176565, Sidman, U. et al., Biopolymers 22:547 (1983), and Langer, R. et al., Chem. Tech. 12.98 (1982).

Alternatively, instead of incorporating the TIGR molecule(s) of the composition into polymeric particles, it is possible to entrap these materials in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylellulose or gelatine-microcapsules and poly(methylmethacylate) microcapsules, respectively, or in colloidal drug delivery systems, for example, liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences (1980).

In an alternative embodiment, liposome formulations and methods that permit intracellular uptake of the molecule will be employed. Suitable methods are known in the art, see, for example, Chicz, R.M. et al. (PCT Application WO 20 94/04557), Jaysena, S.D. et al. (PCT Application WO93/12234), Yarosh, D.B. (US.

Patent No. 5,190,762), Callahan, M.V. et al. Patent No. 5,270,052) and Gonzalezro, R.J. (PCT Application 91/05771), all herein incorporated by reference.

Having now generally described the invention, the same will be more readily understood through reference to the following examples which are provided by 25 way of illustration, and are not intended to be limiting of the present invention, unless specified.

EXAMPLE 1 Single Strand Conformational Polymorphism Single strand conformational polymorphism (SSCP) screening is carried out according to the procedure of Hue et aL, The Journal of Investigative Ophthalmology 105.4: 529-632 (1995), herein incorporated by reference. SSCP primers are constructed corresponding to sequences found within the TIGR promoter and two of exons of TIGR. The following primers are constructed: forward primer "Sk-la": GGC TTC CTC TGG AAA C-3' (SEQ ID NO: reverse primer "ca2": TGA AAT CAG CAC ACC AGT AG-3' (SEQ ID NO: forward primer "CA2": GCA CCC ATA CCC CAA TAA TAG-3' (SEQ ID NO: reverse primer AGA GIT CCC CAG AIT TCA CC-3' (SEQ ID NO: forward primer ATC TGG GGA ACT CIT CTC AG-3' (SEQ ID NO: 10); reverse primer r+2(4A2)"1: ACT TGT TGC AGA TAC G-3' (SEQ ID NO: 11); forward primer -Pr- 5'-ACA ACG TAT CTG CAA CAA CTG-3' (SEQ ID Na& 12); reverse primer 5'-TCA CCC ITA ACT GCA GAA CC-3' (SEQ ID NO: 13); forward primer 5'-ITG C*IT CTG CAG ITA AGC C-3' (SEQ ID NO: 14); reverse primer "Pr+2(4A1)": 5'-AGC AGC ACA AGO GCA ATC C-3' (SEQ ID NO,. reverse primer 5'-ACA GGG CTA TAT TCI' GGG-3' (SEQ ID Na. 16); forward primer "KS1X": 5'-CCT GAG ATG CCA GCT GTC C-3' (SEQ ID NO: 17); reverse primer "SK1XX": 5'-CTG AAG CAT TAG AAG CCA AC-3' (SEQ ID NO: 18); forward primer "MKSal": 5'-ACC iTC GAC CAG GCT GCC AG-3' (SEQ ID NC>. 19); reverse primer "SK3" 5'-AGO TIT T CGA CIT CCA G-3' (SEQ ID NO: forward primer 5'-ACA NIT ACT GOC AAG TAT GG-3' (SEQ ID NO: 21); reverse primer "SK6A": 5'-CCT TCT CAG CCT TGC TAC C-3' (SEQ ID NO: 22); forward primer 5'-ACA CCT CAG CAG ATO CTA CC-3' (SEQ ID NO: 23); reverse primer "SKB": 5'-ATG OAT GAC TGA CAT CCC C-3' (SEQ ID NO: 24); forward primer7"1(6": 5'-AAG OAT GAA CAT OCT CAC C-3' (SEQ ID NO: The locations of primers: Sk-la, ca2, CA2, Pr+1, Pr-i, Pr+2(4A2), Pr-2(4A), Pr+3(4A), Pr-3 Pr-3(4A), Pr+2(4Al), and Pr+1(4A) are diagramatically set forth in Figure 4. The location of primers: KS1X, SK1X, Ks~al, SK3, 1(54, SK6A, SK8, and KS6 are diagramatically set forth in Figure S.

Families with a history of POAG in Klamath Falls, Oregon, are screened by SSCP according to the method of Hue et al, 7Te Journal of Investigative Ophthalmology 105.4: 529-632 (1995), herein incorporated by reference). SSCP primers SK-la, ca.2, CA2, Pr+1, Pr-2(4A), Pr-i3(4A), SKDXX, and KS6 detect single strand conformational ~.polymorphisms in this population. An SSCP is detected using SSCP primers Pr+3(4A) and Pr-2(4A). 70 family members of the Klamath Fall, Oregon are screened with these primers and the results are set forth in Table 1.

TABLE 1 Total SSCI'+ SScCp- Glaucoma positive individuals 1 12 12 0 Glaucoma negative individuals 13 0 13 Spouses (glaucoma negative) 16 2 14 %.Lt4b29 6 23 1 glaucoma positive individuals as determined by 101' of greater than 25 mnmHg 2 unidentified glaucoma due to the age of the individual.

A second SSCP is detected using SSCP primers Pr+1 and CA2. 14 family members of the Klamath Fall, Oregon are screened with these primers. A characteristic polymorphism. is found in the 6 affected family members but absent in the 8 unaffected members. A third SSCP is detected using SSCP primers ca2 and sk- Ia. The same 14 family members of the Klamath Fall, Oregon that are screened with Pr+1 and CA2 are screened with ca2 and sk-la primers. A characteristic polymorphism. is found in the 6 affected family members but absent in the 8 unaffected members. A fourth SSCP is detected using SSCP primers 1(56 and SK1XX. 22 family members of the Klamath Fall, Oregon and 10 members of a Portland, Oregon pedigree are screened with these primers. A polymorphism is found in exon 3. The results are as set forth in Table 2.

TABLE 2 Total SSCP+ SSCP- Klamath Fall, Oregon Glaucoma positive individuals 1 3 3 0 25 Glaucoma negative individuals 6 0 6 Others 2 13 6 7 Portland, Oregon ~i~ *Glaucoma positiveinvdul 6 Glaucoma negative individuals 4 0 4 Others 2 0 0 0 1 glaucoma positive individuals as determined by IOP of greater than 25 mmHg 2 unidentified glaucoma due to the age of the individual.

EXAMPLE 2 TIGR Homologies A novel myosin-like acidic protein termed myocilin is expressed predominantly in Lhe phoioreceptor cells of retina and is localized particularly in the rootlet and basal body of connecting cilium (Kubota et al., Genomics 41: 360-369 (1997), herein incorporated by reference). The myocilin gene is mapped to human chromosome Iq23-q24. The coding region of myocilin is 100 percent homologous with TIGR.

Homology searches are performed by GCG (Genetics Computer Group, Madison, WI) and include the GenBank, EMBL, Swiss-Prot databases and EST analysis. Using the Blast search, the best fits are found with a stretch of 177 amino acids in the carboxy terminals for an extracellular mucus protein of the olfactory, olfactomedin and three olfactomedin-like species. The 15 alignment presented in Figure 6 shows the TIGR homology (SEQ ID NO. 27) to an expressed sequence tag (EST) sequence from human brain S(ym08hl2.rl)(SEQ ID NO. 28)(The WashU-Merck EST Project, 1995); the Z domain of olfactomedin-related glycoprotein from the rat brain (1B426bAMZ)(SEQ ID NO. 29)(Danielson et al., Journal of Neuroscience Research 38: 468-478 (1994), herein incorporated by reference) and the olfactomedin from olfactory tissue of bullfrogs (ranofm) (SEQ ID NO.

30)(Yokoe and Anholt, Proc. Natl. Acad. Sci. 90:4655-4659 (1993), herein incorporated by reference; Snyder and Anholt, Biochemistry 30: 9143-915 3 (1991), herein incorporated by reference). These domains share very similar amino acid positions as depicted in the consensus homology of Figure 6 (SEQ ID NO. 31), with the exception being the truncated human clone in which the position with respect to its full length sequence has not been established. No significant homology is found for the amino termini of these molecules.

While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principals of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains and as may be applied to the essential features herein before set forth and as follows in the scope of the appended claims.

Throughout this specification the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.

9 0 51a SEQUENCE LISTING GENERAL INFORMATION: APPLICANT: The Regents of the University of California (ii) TITLE OF INVENTION: METHODS FOR THE DIAGNOSIS, PROGNOSIS AND TREATMENT OF GLAUCOMA AND RELATED DI SOPDRS (iii) NUMBER OF SEQUENCES: 32 (iv) CORRESPONDENCE ADDRESS: ADDRESSEE: Howrey Simon STREET: 1299 Pennsylvania Avenue, N.W.

CITY: Washington STATE: DC COUNTRY: U.S.A.

ZIP: 20004 COMPUTER READABLE.FORM: MEDIUM TYPE: Diskette COMPUTER: IBM Compatible OPERATING SYSTEM: Windows SOFTWARE: FastSEQ for Windows Version (vi) CURRENT APPLICATION DATA: APPLICATION NUMBER: FILING DATE:

CLASSIFICATION:

(vii) PRIOR APPLICATION DATA: APPLICATION NUMBER: 08/791.154 FILING DATE: 28-JAN-1997 (viii) ATTORNEY/AGENT INFORMATION: NAME: Marsh, David R REGISTRATION NUMBER: 41,408 REFERENCE/DOCKET NUMBER: 07425-0054 (ix) TELECOMMUNICATION INFORMATION: TELEPHONE: 202 383-6904 TELEFAX: 202 383-6610

TELEX:

INFORMATION FOR SEQ ID NO:l: SEQUENCE CHARACTERISTICS: LENGTH: 5300 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ, ID NO:1: ATCZlw'G=T AGTTTACCTC AGGGCTATTA TCCTATAAAC TGTATAGCCT CCATTCGGAT GGAAGAAGGA GTATCCACGT TAGCCAAGTIG CAGATGTTGC TCCTGACAGA AGCTATT7 CATCAAACAG GAGCTAAGAA ACAGGAATGA L'*t i~L~lL.

ATGAGGACCA AAATCAATGA ATAAGGAAA TAATTAAGTA T7T=T'CTT GGGAAGAGAC AAACGTCAAA AGCATGATCT GATCAQATCC

TGAAATQAAA

GTATGTC=~

T CCAGGCTG

CAGGAAACAT

CATGGvGCACT TCc 7-7

CAGCTCAGAA

CTCCATG=A

CAAA1GTCGAT

GGTC-ATGTA

GAA.AATGGGG

ACCTCCCCGT

GAAATATGAA

GAGCCTTCAA

CCAG IWTTC GGCATCACTC TGGCGAGGCA AGCAAAATCA AAATTCCGCA A5GTGA7TAGG CAGTTGACCA ATM'ACTGGG CTAAGCCTIGG GACATGGTTA AAAGGCAACC GGGACCCTGA GGCATWIGCC TTGAAAGATC ATC.AATTTTA T1TAGACATG GGTCCCAATT GGATAGTCA GAAATCATTA TGTCATAGCC CTCACACACA GTGCCTCAAC CATTGTTAAC TGTGCAGCCC ATCCCGCTCC TACAGCCGA AGCTCCGTGA ACCTGAGCTC ACTGCAACCT CGCGTAGCTG GGACTACAGG GPTrCACCAT ATTAGCCCGG AGCCTCCTAA AGTCTGGGA TTAATAAGGA ATAACT1TGAA TAATr'rCAGG GATTC7rG

AGTICAGGAA

AATGCAGGAG

TGT7CGCAAC

ACTI'TCAADG

AGAACAT1'GT

=~AGGAAGG

ACCATTAA GTATAAAACA 7TATAAAAGTC AGGCATACAA GAATACTIG T=CCCATC GGCCCGATGT GT CTGACCTA GTGTICATCTC AGTAGGTCCC ACAGCGAAGTC TVCCCACTCT GGGTGAGGGT CTGTGTCTTA CTGCCTCCCA CGTCAGC

CACTGGTCT

CACCAT'GCWI

TTCCATTG

T3AACCCCC T ,GCGTCCT GACCTGT1'GC TAT1'GAGTAC

CCTACCTTCG

CATCACTTTC

~T GTAAG

GGCCMTCG

GGCAGAGG"T

GAGCAACC'G

1-rTCTATTTC 7TATATCTGC

TGGAGGTGAC

CGCACGCCCG

CTGCTG

TTACAGGCAT

TGGTTTACTA

ATGGGGAATG

'rrCCTCATC

CCTCCACATC

GTCTCTQGTAT

TCCT CT CCAG

CCAGCCCGTG

TGV7ACTC

CAGACACCASG

IflTTTCTC-AT

GAAAGAAAGG

CCT CCCTAGC

AAGCCTCCTC

GAAACTAGTC

CACGTCCAGG

GCTAATTTT

ACTCCTGACC

GAGTCACCGC

AACCAACAG

G7CCCATGAG CTCAT~rTCA G7TACTAAA

AGGGGAGGAG

CTGGGGGAGC

CCACTGGQTlG

GTTCATTCAT

AGACAAAATG

GGAAGACGTOG

AAATAAACAC

GCCCCCT=C

CTCCATCACT

TAACGGAGAA

AGAATTCCAG

TGxAGATAACC

GGCAGGATGA

G~t~l=

CACATICCAAT

TGCCCAAGGA

AAAAGATGT=

GCTT1ATGGG

TATTATI'TTA

GCAAAGGACA

ACTOGGAAAG

CTATACCAGG

AAACTGAGAG

AGCAGCAGTG

TAAGGAATCT

AATATrjCGAT GGATAAOGTr.

CTAACTr

CAACCACATC

ATTACAAATG

AGACTYICTGC

CACCTACCTIG

AT7CTCCTGT

GTATIYGPAG

7rCAGGTGATC

GCCCGGCCAA

GAAACA%2ACA

CTGCCTGCCT

GGCTAAG=rA

TAAGAGTATA

GGCATACCCC

CCTG.CAAGCA

T7TrGTAT'C CCAGGCMArC

GTGAGCAAAC

CAGAGAAGA

GCCTCCATCG

CACAGCGCTG

AACTCCT

TCTGGAGGGG

GAGGTGGGA

GTr&AAAAGC TCMAAGGGv

TCCTTCAOGA

TAATTCAG

177CTTTCTGG

GGGCCATAGT

CTG7GATTCT

OCCACAQGG

TCACTGATCA

CCCAACArvCC GAATCAGAGv GTGvGAAAATC

AATGTGMAAAG

TAAAGAM'CA

ATTTTAG7GA

ATGGTAAATC

AAAATGCCAG

TCCAAATG

AAAATG-GGAA

AAAACCAGAT

TAACAATAAC

CTTCATAAC

GAACACAAAA

CAAAACAAAAA

CCCCTCAkCA

TAAGAAACTC

GCATAATCAG

TCCCAGCTCC

CAGAATGATC

TACAACCCAA

CCACCTCCCC

ATCACGATGT

TATGCTCTAC

CTCAGCCTCC

TAGAGATGGG

CACCCACCTC

GG'TCACT'GT

AAAGCTGTGA

A=JCCCAGAC

CATTTTATT

CATAAACTAG

AGAGACTCCT

CCCGGGGTCC

ACTCTCTAGG

ATTGACAATT

CAGTCACTGC

TTAATAGCCA

AA7TCGC

TGCCCCGAGG

CAGCTG.GCCT

GCTCCAGGCT

ACAkGTGTTC

CTGCAGGGAG

CAAGGCTGAA

TCCTGTGAGC

ATGCAGTIVIC

AGGCCTTTAT

AAGAAGTIGAC

ACTT~CC='A

CTTAGGG

AGGTGGAGGG

CGTCAGACTIC

ACATTTr=C ACTAAC7GQT

CCAGAGATTG

120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 31iso 3240 3300 3360 GCCAAC7ZWAA ACCCAGTGCT GCA7CCC7 AACAC=CA CCCCCAAGCC CGAGTCM1CC CTCAGAGGGA AAGGGGCCTC GGGACGCT GGCGQC QGGTCTrAA AGQCAGGAAG GCTGvCCCAGA TGTTATGT :CIAOGGG GCTGQAGTT CTTTTATCT TCGAGGAGAA GAGTCTATT ATAAACTCADG CTGTTAAAAT T CGGG CoCATGGGT'1T TrAATGGGAA TATA1GQAAGC GAGCTCATrP TGQAGTCn'r TGCAAG.AcGG Gl#.GCjTCT

ATAAAGACCC

GA7GTT=A

G=CAGGCAAG

CAGGACCGAG

TCCCTAAGCA

GGTAGC2TI TCTITrCATGT

TCGAAAACCT

GCGACCGTGG

AGGGGAAGGA

GCAGGCAGAA

AGCCACAATG

TAGACAATGG

GCCTr.CATT

GCAATC

GGCAGAGCTG

CAT17GCCAA

CAAAAACTGG

CCTAGGCCGT

AGACGT

GAACACTTCC

CAGCAGCTGA

CCA7CATC

GCTCAATGAA

TAACCAAAAA

CCCAGACCAA

TTAA.ACTTTT

AGTGACCTGC

ACAGATTCAT

GTTCTAGGAG

CCTGA171CT

GTAGTAACTG

GGAG2'rAGCA

TAAAGCCAAA

CAT7 TCP.GCG

TAAACAAACA

AAGAATAGAA

AGTTTGGAAA

TOCTAAAGG

TTATTGGCTA

GGATTAITTAA

NTT~TTACC

CTCAAAGTGG

TTATACTATA

CmGAATI' ATI1 'GATAT

ATATATI'TGA

CMTGCACACA

TGCAAGACTG

AGGGGGGAAA

A 'JA.JT TC.

GGATCTCCAG

TC.AATGGAAA

TGTAAGTGTG

TAGGAACTAT

CAAACAGACT

CCT GTGCACA

GCTCCCCACT

CAGGCACCTIC

CACCCTGACC

AGCGCAGGGG

TCAAGGGCAG

GCAGGGCTAT

AATACTATAT

AGGCTGTAAG

GCACAAGGGC

CAGATTCAAG

ATGTTNACTA

ccc~qIj p?

TCTTAGAGC

TATTTACTTC

CAATCATrTAT

T'IGCCTTT

CCTACAGTCC

ACCTTICTAAC

TAATAACAGT

TTACAGTTGT

AGACCTCZCTG

TTTGATAATC

AAACACTT

CACAGAGTAA

AAATTAGAAA

TCTGCCGCTT

GCTA.GGCTGJA

TTCCTAGCAT

TATAAACTAG

TGTACGTGTG

TA77'GGGGTA

TCTGGAAGGT

GCCCCACCCA

ATATATAAAC

TCXGCAC.AGC

AGCACCCCAC

AGGAGAAGAA

TGGGAATA

ATTGTGGGGG

TTTCCTA

ATTACTTAGT

AATCCCCTT

CCTAGGTCTT

TCTTCAG

AXACTCT=

ACAAGTATrG 77CAAGTGQC

AGAAAGCCTIG

TAAATTrAAC ACCTGTGATr

TCCAGATACG

CTGGATZTTG

ATATr TCATT

CTGAGAAGAG

GAACTGATr

GTTCTCCCAA

CTATAGGAAT

TATTIT1'CTC

AGTG;CCTGGC

AAATATATCC

TGGGT13CATA

TAVTTTICTAA

GCCTCACGTG

CTCTCTGOGAG

GCAGCTCAGC

AAAGAGAGGG

CCACAGGGAT

GAAAAAATCA

CAAGCTGAGT

TI'CTCCTrTAT C71T 7AACAG

GCTGACTATA

AAAATGAGAC

TCTCCACTCT

ACACTGTI'GT

TTAAAI2TTAC TGsAATTTGAA

ATTTATTCC

77GCATAC

TTGTAAGTGA

T1TTTTAACAT ATCATTr

TI'CCCCAGAT

AGAGGCTAAC

AAGATACACAG

GCTC TCCCTG

TGTCCCTGCT

ACAfGTrCAGG

TTUTTCGAAAT

ATa TD Aa AGP A

AATTGGATG

GAATT~rCT

GCCACCTCTG

CTCGGGCATG

AGTCACTGCT

ATAGTGTATG

TATAGTCCAC

GTTAAGGGA

AATI'CTGAGC

TAGGAACTCT

GAAGAAAACA

'I~GG~

TAGTACCCTT

iAL1T

CCAGGTGAGT

TCGTATTAAC

TTCTGACAGT

GGTTT~ATTAA

ATTGCGAATA

CAATAGAAAT

AATATTTATA

ATTAATAAAA

TCCTTGTAA

TTCACCAATG

AT 'GACAT'rG T7GTTTTAAA

GAGCCTGGTA

ACGTCTTAAA

TTCTCAATGA

CAGCACACCA

TGTAAAACCA

TTCl 7TAA

GGCAGCGTGA

TCTTCCCCCA

GACAGCACGC

AGCAAGAAAG

G7K:ATCCTCG

AGTCGGGAGA

AAGTCACAAG

T1'CCTAAGAG 777AAAAAT

CCAATCAAAT

C.TGCCAGGGC

AACATAAAGT

1I'GTATAT

TGTAAAGCAG

ATTACArTr

GAGCCATAAA

CACAGACA7T

CTCAAAACTA,

CATGTTAAA

TCTATATTTr

AGGTTCTTGG

GTGCCTrGAGA GCTAG3GGGTrG

GGACTTTT

GTTTGCAGAG

GTAGTCCTGG

GGTGGAGATA

AAAGAAACTC

AGGCAACCCC

TGAAGGGCTG

3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 5300 AGCCAGCAAG GCCACCCATC INFORMATION FOR SEQ ID NO:2: SEQUENCE CHARACTERISTICS: LENGTH: 5304 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID N0:2: ATCTTTGTTC AGTXTACCTC AGGGCTAT'A

TCCTATAAAC

GGAAGAAGGA

CAGATGTTrGC

CATCAAACAG

GAGAGCAAAT

ATGAGGACCA

TA7ATZAAGTA

AAGTVAAA

GGCATCACTC

AGCAAAATCA

AGTGATTAG

ATTGACTGGG

GACATGGTTA

GGGACCCTGA

TTATAGCCT

GTATCCACGT

TCCTGACAGA

CAGCTAAGAA

AATGATGAAA

AAATCaATGA

AGCATGATCT

TGGGGAGGCA

AAA7CGCA

CAGTTGACCA

CCAT7V=GT

TAGCCAAGTG

AGCTATTCT

ACAGGAATGA

AATAAAC7T

ATAAGGAAAA

GGGAAGAGAC

GATCAGATICC

AGT7 CAGGAA

AATGCAGGAG

r.T~CAAC

TGOAA.ATGAAA

GTA7KGTCTT

TCCAGGCTGT

CAkGGAAACAT

GATGGGCACT

CAGCTCAGAA

TGAGATAACC

GGCAGGATGA

CACATCCAAT

TGCCCAAG2GA TTTAATrT!CA

AAAAGAGT

CCCATGTIGA CCTATGG CAAAGTGGAT TATTATITTA GGTCATGTTA GCAAAGGACA GAAAA1GGG ACTGGGAAAG ACCTCCCCGT CTATACCAGG GAAATATGAA AAACTGAGAG GAGCTCA AGCAGCAGTG CCAGTI'ITCT TAAGGAATCT

AATGTGAAAG

TAAAGAATCA

ATTTTAGTGA

ATCGTAAATC

AAAATUCCAG

GGAAAAAATG

TCCAAATG

AAAATGGGAA

AAAACCAGAT

TAACAATAAC

CTTTCATAAC

GAACACAAAA

CAAAACAAAA

CCCCTCAUCA

TAAGAAACTC

CTAAGCCTGG ACTTTCAAGG AAAGGCAACC AGJAACATTGT GGCATTT1GCC TTTAGGAAG IlrAACATC 1TTTAGACATG GGATAGGTrCA 7UTCATAGCC

GTGCCTCAAC

TG7GCAGvCCC

TACAGCCAGA

ACC7GAGCTC

CGCGTAGCTG

G =11LACCAW

AGCCTCCTAA

TTAATAACGA

TAA777rCkGG

CACTGGTCCT

CACCA7GCT

TTCCATN'G

TGAAGCCCCC

TCGGG1=CT GACC7TTGC TAT1'GA=TAC

CCTACCT?'CG

GCCAACTAA

AGCATCCCTT

CCCCCAAGCC

GCC7CGCTTC

CCAGAAAGGA

CTCA.GAGGGA

7UGGGACGCT

GCTGCCCAGA

CTTTITATCT

ATAAA1GTCAG

TAAGGAA

TGCAAGACGG

GGTIJL'GCLLGT

ATAAAC4CCC

GGATCTI'GAG

GGACAGGAAG

CAGGACCGA

TICCCTAAGCA

GGTAGCTrr

TTAAACTT

AQTrAACCTGC ACAaATICAT

GTTCTAGGAG

CCTraAmT GTACTAACTro GGAG7TACA TAAAGCCAA6A CA77CGCM

TAAACAAACA

AAGAATAGAA

AGTTTGGAAA

TGCTCAAAGG

TTATrioGCTA GGATTATrAA TAT1Tfl=ACC CTCAAkAG=G

TTATACTATA

ClTrTrAAATT Alr.AATTTA GGTCCCAATr

GAAATCATTA

CTCACACACA

CATTGT1AAC

ATCCCGCTCC

AGCTCCGTKGA

ACTGCAACCT

GGACTACAG

AGTG.CTGCGA

ATAACTTGAA

GATCTGG

CATCACTTrC

TTGTCGTAAG

GGCCATC7TGT

GGCAGAGGTT

GAGCAACCTG

7TATATCTC 7rGGAGGTGAC

ACCCAGTGCT

AACAAGGCCA

CGAGTCTTCC

CCCTr.AATCG

AATGGAGAGG

AAGGGGCCTC

GGGGCTGAGC

TiGTTCAGTGT

CTGTIAAAAT

TATAGGAAGC

TCTTTATCT

TCGAAAACCT

GCGACCGTGG

?TCA-GCTCT

AGGGGAAGGA

GCAGGCAGAA

AGCCACAATG

TAGACAATGG

GCCTGQCATr

CACCCTGACC

AGCGCAkGGGG

TCAAGGGCAG

GCAGGGCTAT

AATACTATAT

AGGCTGTAAC

GCACAAGGGC

CAGA1CAAG

ATG=IACTA

CCCATIGTA

TITAGAGC

TAITTACTIC

CAATCATTAT

TTCCATTT

CCTACAGTCC

ACCTTCTAAC

TAATAAGAGT

TTACAGTTGT

AACCTC=T

ACCATTTITAA

TTATAAAGTC

GAAATCACTG

GGCCCGATG

GT3TCTCT

ACAGGAA=T

GGGTCAGGGT

CGCACGCCCG

T1TACAGGCAT

TGGTTTACTA

ATGGGG-AATG

TT'CCCTCATC

CCTCCACATC

GTGTC=GAT

TCCTCTCCAG

CCACCCGTG

7TGCTACTC

CAGACACCAG

AGTITCTCAT

GAAAGAAAG

CCT'CCCTAGC

AAGCCT'CCTC

GAAACTAGTC

CACGTCCAGG

GGG TGCTGAA

ACGG

TCCXGCGTGT

GAGCTCATIT

TGGAATCAGG

GCAAGT

GGCAGAGCTG;

CA7TTGCCAA

CAAAAACTGG

ACCACCCCAC

AGGAGAAGAA

TGGGAAT1IGA A7TGQGQ T777WCC7TA ATTACTrACT

AATCCCGTWP

CCTAGGCT

TCTCATTCAG

AA1CCTAA

AAACT=

ACAAGTAT1'Q

TTCAAGTGGC

-C S S JGJS A

AGAAAGCCTG

TAAATTTAAC

ACCTGTGA1IT

TCCAGC;TACG

CTGMTCT1r.

GTATAAAACA

AGGCATACAA

TGTCCCCATC

GTCTGACCTA

AGTAGGTCCC

TCCCCACTCT

GCTAATT~1rT

ACTCCTGACC

GAGTCACCC

AACCAACAGG

GTGCCATCAG

CTCATrCA

GTACTGAAA

AGGGGAGGAG

C7GGGGGAC CCACTGvGTM' GT1'CATTCAT

AGACAXAAATC

GGAAGACGTG

AAATAAACAC

GCCCCCTGCT

CTCCATICAGT

TCTGAGCTG

TAACGGAGAA

AGAA7TCCAG

AGGCAGGAAG

GCTGGGAGTT

GA.GTCTATT

GATGGGTrr

CCTAGGCCGT

ACTACTCAC

AGACTCGGZT

CTCCTTCCCT

GAACACTICC

GAGCAGCTGA

CCATCACTCC

GCTCAATGAA

TAACCAAAAA

GCCAGACCAA

GCAGCTCAGC

AAAGAGAGGG

CCACAGGGAT

AATATC=CT

GCATAACGTQG

CTAACTTT

CAACCACATC

ATTACAAATG

AGACTTCTGC

CACCTACCT(3 ArrCTCCTGTT GTAT7=TAG

TCAGGTGATC

GCCCGGCCAA

GAAACAGACA

CTIWCTIGCCT

GGCTAATA

TAAGAGTATA

GGCATACCCC

CCTGCAAGCA

T1?I'G=ATC CCAGGCA17C

CACAAGAAAA

CA1tTTAAG

GCCTCG

CACAGCGCTG

AGACTCCTTrG Ttlr.. GG GAGGTGvGGGA

G'C.AAAGGG

ITCCGTTGCT

TCATr.AAGW TCCT~1WCA

TAATTCACGG

c.ciG*%rGMG TTC77TS S.IZTGG

GGCCCATACT

CT=TICT

GCCACAGGGG

TCACTGATCA

CCCAACAGCC

GAATGCAGAG

GTGGAAAATG

AGTGACTGCT

ATAGTGTAT G

TATAGTCCAC

GCATAATCAG

TCCCAGCTCC

CAGAATGATIC

TACAACCCAA

CCACCTCCCC

ATCACGATGT

TATCCTCTAC

CTCAGCCTCC

TAGAGATGG

CACCCACCTC

GGGTCAG=C

AAAGCTGTGA

AGTCCCAGAC

CCATTPATT

CATAAACTAG

AGAGACTCCT

CCCGGGGTCC

ACTCTCTAGG

ATTrGACAA'rT

CAGTCACTGC

TAATAGCCA

AATTGTIXCGC

TGCCCGGAGG

CAGCTGGCC

GCTCCAGGCT

ACAGTGT=T

CTCAGGGAG

CAAGGCTGAA

7TC1AGC ATGCAGT1TC

AGGCC'TTAT

AAGAAGTGAC

AC7TGC A CTGTCGZxrmG

CGTCAGCGI

ACATmrrVCT

ACTAACTGGT

CCAGAGM'1'G

GACAGCACCG

GTGATCCG

AGTCGGGAGA

AAGTCACAAG

7777t"IrTG1TJ

TTCCTAAGAG

1?I'GAAAAAT

TUGTCAGCTG.

CCAPLTCAAAT

C7CCAGGGC

AACATAAACT

TTT=GATAT

TCTAAAGCAG

ATTACGTrT

GAGCCATAAA

CACAGACA7T

CTCAAAACTA

CATGM~AAA

960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 GAAAAAATCA GTTCAAGGGA C-ACCTGAGT AATCGC TTCTCCM AT TAGGAACTCT CT1TrAACAG GAAGAAAPACA

GCTGACTATA

AAAATGAGAC

GTT..XGCj.

TCTCCACTCT

ACACra=

TTAAAGTTAC

7CAArAA A1?1'ATTCC

ATTAC

TI=AAGTQA

1?rTACAT

TMCATVPTT

TAGTACCCTT

AACTGCAQ.AA

GGAGGTGAGT

TGQATrAAC

TTCTIGACACT

001 TATTAA

TGGGAAA

CAATAGAAAT

AATATATA

ATTAATAAAA

ATTTGATAT TITGATAATC ATATATTXrGA AAACATCTTT CATISCACACA CACAGAGTAA TGCAAGACTIG AAATTAGAAA GGATCTCCAG TTCCTAGCAT TGAATGGAAA TATAAACTAG TGTAAGTGTG TGTACGTGTG3 GATATAGGAA CTATTATTIGG ACTCCAAACA GAZIT7TGC"Ar CCCCCCT13TG CACAGCCCCA GCTGGCTCCC CAGTATATAT CATCCAGGCA CCTCTCAGCA

ATATTTCATT

GAACT3AT

GTTCTCCCAA

CTATAGGAAT

TATI'rrrCTC

AGTGCCTGGC

AAATATATCT

13 TGTGTGTG

GGTATGGGTG

AG.GrATTr

CCCAGCCTCA.

AAACCTCTCT

CAGC

ATCATTTGTT

TI'CCCAGAT

AGAGGCTAAC

AGATACACAG

G.CTC TCCCTG.

TGTCCCTGCT

ACAGTGCAGG

0GTIAAAT

CATAAATTGG

CTAAGAATCT

CGTGGCCACC!

GGAGCTCGGG

TCCIVIGTAA TCTATATTTT

TTCACCAATG

ATTGACAro IUG 'IAAA

GAGCCTGGTA

ACGTCTTAAA

TTCTCAATGA

CAGCACACCA

TGTGTGTAAA

TGCTGGCAGC

TCTGTCTwrCC

CATGAGCCAG

AGGTTCTTGG

GTGCCTGAGA

GCTAGGGGTG

GOGTGCTGTC

GGACT~GT=

GTITGCAGAG

GTAGTCCTGG

ACCAGGTGGA

GTGAAGGCAA

CCCATGAAGG

CAAGGCCACC

4560 4620 4680 4740 4800 4860 4920 4980 5040 10 1.,A 5160 5220 5280 5304 INFORMATION FOR SEQ ID NO:3: SEQUENCE CHARACTERISTICS: LENGTH: 6169 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:

ATCT'I'TGTTC

TCCTATAAAC

GGAAGAAGGA

CAGATGT~rGC

CATCAAXACAG

GAGAGCAAAT

ATGAGGACCA

TAATTAAGTA

AAACGTCAAA

GGCATCACTIC

AGCAAAATCA

AGTGATTAGG

ATTGACTGGG

AGTTTACCTC

TGTATAGCCT

GTATCCACGT

TCCTGACAGA

GAGCTAAGAA

AATGATGAA

AAATCAATGA

i'rGTCCT'

AGCATGATCT

TGGGGAGGCA

AAATrCCGCA

CAGTI'GACCA

CTAAGCCTGG

AGGGCTATTA

CCAWGGAT.

TAGCCAAGTG

AGCTAT'rCT

ACAGGAATGA

AATAAACTTT

ATAAGGAAAA

GGGAAGAGAC

GATCAGATCC

AGTTCAGGAA

AATGCAGGAG

TGT TCGCAAC AC7rTCAAGG

AGAACATIT

TTTAGGAAGG

ACCATTTTAA

TTATAAAGTC

GAAATCACTG

GGCCCGATGT

GTGTCATCTC

ACAGGAAGTC

GGGTGAGGGT

TGAAATGAAA TGAGATAACC GTATGTCTI GGCAGGATGA TCCAGGCTGT GCGTT CAGGAAACAT CACATCCAAT GATGGGCACT TGCCCAAGGA TCCCTTGTT TTTAATTTCA CAGCTCAGAA AAAAATGT CTCCTGTA GCTTGATGG GACATGGTTA AAAGGCAACC GGGACCCTGA GGCATTTG;CC TTGAAACATC ATGAATI'rrA AGACATIG GGTCCCAATT GGATAGGTCA GAAATCATTA 2GTATAGCC CTCACACACA *GTGCCTCAAC CATTGTTAAC TGTGCAGCCC ATCCCGCTCC TACAGCCAGA ACCCTG ACCTGACTC ACTGCAACCT -CGCGTAGCTG GGACTACAaG GTI'TCACCAT ATTAGCCcGG AGCCTCCTAA AGTGCTGGG TTAATAArGGA ATAACIGAA TAATITrCAGG GATTCTT GG CAC TGGTCCT CATCACTTTC CACCATGCTIT TI'GTGGTAAG L777W~ GGCCATCTGT TGAAGCCCCC GGCAGAGGTT Ta9GG1TCT GAGCAACCTC GACCTGTrGC TTTCTAT?1'C

CAAAGTGGAT

GGTCATGTTA

GAAAkATG4GG

ACCTCCCCGT

GAAATAT'GAA

GAGCC WICAA

CCAGTTC

GTATAAAACA

AGGCATACAA

TGTCCCCATC

GTCTGACCTA

AGTAGGTCCC

TCCCCACTCT

TA~wrATTTTwA

GCAAAGGCA

ACTGGGAAAG

CTATACCACG

AAACTGAGAG

AGCAGCAGTG

TAAGGAATCT

AATATGCGAT

GGATAACGTG

CTAACTTvrr1T

CAACCACATC

ATTACAA.ATG

AGACTCG

CACCTACCTG

ATTCTCCTGT

GTATTGTTAG

TCAGGTGATC

GCCGGCCAA

GAAACAGACA

CTGCCTGCCT

GGCTAAGTrA

TAAGAGTATA

GGCATACCCC

CCTGCAAGCA

T1SI1WI=TATC

CCAGGCATTC

AATGTCAAAG

TAAAGAATCA

ATTTTAGTGA

ATGGTAAATC

AAAATGCCAG

GGAAAAAATG

TCCAAATTGG

AAAATGGGAA

AAAACCAGAT

TAACAATAAC

CTP1'CATAAC

GAACACAAAA

CAAAACAAAA

CCCCTCAGCA

TAAGAAACTC

GCATAATCAG

TCCC.AGCTCC

CAGAATGATC

TACAACCCAA

CCACCTCCCC

ATCACGATGT

TATGCTCTAC

CTZAGCCTCC

TAGAGATGGG

CACCCACCTC

GGGTCAGTGT

AAAGCTGTGA

AGTCCCAGAC

CCATTTTATT

CATAAACTAG

AGAGACTCCT

CCCGGGGTCC

ACTCTCTAGG

ATTGACAALTT

120 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 CGCACGCCCGG GCTAATT~wZT LAUL I~ ACTCCTGACC TTACAGCAT GAG ICACCGC .GTIACTA AACCAACAOG ATGGGGAATG GTGCCATGAG TICCCTCATC CTCATrr1CA CCTCCACATC GTTACTGAAA CTGTQTGTA AGOGGAGCAG TCCTCTCCAG CTGGGGGAOC CCAGCCcGTG CCACTGGTTG TGTGTGCTC GTTCAEITCAT 56

TATTGAGTAC

CCTACCTICG

GCCAAC7TAA

AGCATCCCTT

CCCCCAAGCC

CTCAGAGGGA

TQGQACGCT

GCTGCCCAGA

C.777TrATCT

ATAAAGTCAG

TAATCGGAA

I ;AGTCT~r

TGCAAGACGG

CvL77GCG...-

ATAAAGACCC

GGA7TTGAG

GGACAGGAAG

CAGGACCGAG

TCCCTAAGCA

GGTA~cTrrrT TT1AAACTr

AGTGACCTGC

ACAGATTCAT

GTTCTAGGAG

CCTGATTrCT

GTAGTAACTG

GGAG.ITAGCA

TAAAGCCAAA

CAT1W7CAGCG

*TAAACAAACA

A1GAATGAAT AG7 TTQGAAA

TUCTCAAAGG

TrATT'GGCTA GATTA7TAA TG7TTwZTACC

CTIZAA.AGTGG

TTATACTATA

CTT 'GAAA1T ATmT1'GATAT

ATATATITGCA

CATGCACACA

TGCAAGACTG

AGGGGGGAAA

C77GTGTTICTA=

GG.ATCTCCAG

TC.AATGGAAA

TGTAAG1W

TAGGAACTAT

CAAACAGACT

CCUTGCA

GCTCCCCACT

CAGGCACCTC

ACCAGAGTGG

GCTTAC.ACCT

7TATATCTGaC

T.GAGGAC

ACCCAGTCC

AACAAGGCCA

CGAGTCTTICC

CCGTGAATICG

AA7GGAGAGG

AAGGGGCCTC

GGGGCTGAGC

7CAGTGT 711rCCT CM 7TAAAAT

TATAGGAAGC

TCT A=G

TCGAAAACCT

GCGACCGTGG

7*CAGCTCT

AGGGGAAGGA

GCAGGCAGAA

AGCCACAATC

TAGACAATGG

GCCTGGCATT

CACCCTGACC

AGCGCAGGGG

7CAAGGGCAG

GCAGGGCTAT

AATACTATAT

AGGCTGAAG

GCACAAGGOC

CAGATTCAAG

ATG=IACTA

CCCAGTIGTA

C1'rAGAGCA TATTTAC1'TC

CAATCATPAT

TTGCCATTM

CCTACAGTCC

ACCTTCTAAC

TAATAACAGT

TACAGT1'GT

AGACCTCCTG

TrlrxATAATC

AAACTCTT

CACAGAGTAA

AAA7TAGAAA T'cTGCAT

TATAAACTAG

TATI'GQGTA

GCCOCACCCA

ATATATAAAC

TC-AGCACAGC

TGCGTTGC

CCGATGCCAG

CCAGGCCATC

GGAGGCCACC

CAGACACCAG

AGTIWICTCAT

CCTCCCTAGC

AAGCCTCCTC

GAAACTAGTV

CACGTCCAGG

TGTCGCGG

TGGAGGAGAA

TCCAGGGTGT

GAGCTYCA 'IT LA A.CTGGC

TGGATAGG

GCAAGG=

CGCAGTkCTC

GCGCGGCT

CTTCAGGAAA

CATTCCAA

CAAAAACTGG

AGCACCCCAC

AGGAGAAGAA

TGGGAATTGA

ATTACTTAGT

AATCCCGTT

CCTAGGTCTr

TCTGATTCAG

AAG7TA AACrG1*

ACAAGTATTG

7rCAAGTGGC CAA AA'.IAA

AGAAACCCYG

TAAATITAAC

ACCTGTGATT

7GCAGATAC L'CGATCTrG

ATATTTCATT

CTGAGAA=A

GAACTGATTT

AAA~

CTATAGGAAT

AGACAAAATG

GGAAGACGTG

AAATAAACAC

GCCCCCTGC'

~CTCAT

TC7AGTG

TAACGGAGAA

AGAATTCCAG

AQCCAGGAAG

CTGGGAGTT

CAAGTCTATr

GCATGGGTTT

CCTAGGCCGT

ACTACTICAGC

AGACT:GGT

CTCCTTCCCT

GAACACTTCC

GAGCAGCTGxA

CCATCAGTCC

GCTCAATG.AA

TAACCAAAAA

GCCAGAGCAA

GCAGCTCAGC

AAAGAGAGGG

CCACAGGGAT

GAAAAAATCA

CAAGCTGAGT

TZ'CTCCAT

CTTTAACAG

GCTGACTATA

AAAATGAGAC

G77rCkGGCT

CTCCACATCT

ACAL"ITPGT

TTAAAG'rrAC T77CTCT

T=ATTNGAA

ATTTTAT=C

TT'GTCATTAC

TTGTAAGT'GA

TTTflTAACAT

ATCATT=

CTC.AGCAAAG

CATCTTGAAG

CCCTICCATCG

CACAGCGCTG

AGACTCCTK

GAGCTGGGGA

TCAGAAGGG

TCCTTACGA

TAATTCACGG

CCTGTGGTGG

TTCIT1'CTG

GGGCCATAGT

CTGTATCT

GCCACAGGGG

TCACTGATCA

CCCAACAGCC

GAA7GCACLAG GrcGAAAATG

AGTGACTGCT

ATAILTGTATG

TATAGTCCAC

AA?1TAGC

TAGGAACTCT

CAGTC-ACTC

TAATAGCCA

AATUTCGC

7CCCGAGG

CACCTGGCCT

GCTCCAGGCT

ACAG TT C7GCAGGGAG

CAACCCTIIA

TCC1GC

ATGCAGTTTC

AGGCCT7TAT

AAGAAGTGAC

ACT1'GCCTTA Tii;CCAT LA CA

CTGAGGG

AGGCAGGG

CGTCAGACTC

ACATTICT

ACTAACTGGT

CCAGAGATTG

GACAGCACGG

AGCAAGAAAG

GTGATCCTGG

AG7CGGGAGA

AAGTCACAAG

1TTCTCTGT GAAGAAACAT1CCTAAGAG TGAT'GGT7T771'TGAAAT TAGTACCCTr TGGTCAGCTG AACTGCAGAA CCAATCAAAA QGAGGTGAG CTGCCADGGC TGTATTAAC AACATAAAGT ;1CTACACT TTYGGTATAT GGrTAA TGTAAAGCAG TGAGGAAAAA ATrACATTr ATI'GCAATA GAGCCATAAA CAATAGAAAT CACAGACATr AAT1ATrATA CTCAAAACTA ATrAATAAAA CA7GTTAAA TCCTI 'GTAA TCTATATTTT 2100 2160 2220 2280 2340 2400 2460 2520 2580 2640 2700 2760 2820 2880 2940 3000 3060 3120 3180 3240 3300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 4320 4380 4440 4500 4560 4620 4680 4740 4800 4860 4920 4980 5040 5100 5160 5220 5280 5340 5400 S460 5520 5580 5640 TI'CCCCAGAT TTICACCAALTG AGGT'C~TIGG A.GAGGCTAAC ATTGACATTG GTGCCTCAJ= AGATACACAG TTGTTTAAA GCTAGGGQTG GC'TtTCCCTG GAGCCTGGTA CGGTCCTGTC TATTTT CTCGCT AGTGCCTGGC ACAGTGCAG AAATATATCC TrTTTGAAAT A..DA'.D .U LflwLIWJLWIUT TGQCTGCATA AATIGGGATG TA?1'TTCTAA GAATCTTYGCT GCCCD CCACCTCTG ACGIVTTAAA GGCACTI'Tr1' TTCTCAATGA GTTTGCAGAG C7AGCACACCA GTA*GTCCTGG TGTAAAACCA OGT~rQAGTA 1rI=1~AA AAAGAAACT CTCTC7GGAG

AGACTTC

TGAGCTTTUA A

GAMGGGGW

TATACCT1'CA

TCAGTCAICC

AAAGCTCGAC

CTCGGGCAG

AZAGGAMCC

GGCCTGAGAT

CCAQGACAGC

GTGQCCAG

ATAAC?1TACA

TCAGCT=CC

COSCGGA

A A A A

AGCCAGCAAG

TCACCAAGCC

GCCAGCT

TCAkGCTCAGG

TCCCAATGAA

GAGAGACAGC

GCGAAGCCTC

AGCAACCCC

Tr.AAGG=

GCCACCCATC

TCTrXCAATGA

CAGCTGCTGC

AAGGCCAATG

TCCAGCTGCC

AGCACCCAAC

CTCCACCAAT

TGACCTTGGA

GCACCCTGAG

ACAGCAACCT

AAAATGAGAA

GGGGCCA=T

GTAAGAATGC

CTACAGCoCC

AGATOAATTA

AGCTTICATrr CCAGGCTGcc

GCGGGAGCGG

CCTCCGAGAC

TCTGGCCAGG

TCCCCAGACC

AGAGTGGGGG

TCC.AGGCCTC

AGGAAAGCAC

AGATTAGTGG

AGoGCCCCAGG

GACCAGCTGG

AAGTCAGT

AGGT~rGAAA

CGAGACACTG

GACTCTGAc;T

CCTGCCCT

ACGATCACCT

TTCAGAGTTC

AGACCCAGGA

AAACCCAAAC

TCtG2-r

GCAGCAGCCA

CTGGCG

TCAGCAGGTG

CTCCTAGAGA

TCAAGTATrA

TTGTGCCCCT

GGCGCAG

CAGAGAG7

GAAGAAGCGA

GGAGGTAGCA

GCCACCAGGC

ATATGGCTCG

CTG.CACAGCT

CTAGTAATTr

CCATGTCAG

AGGoGAGCTGG

GAGACTGCCT

CTAAGGCAAC

AGGCTGAGAA

TCCAGAGAAG

TAGTGACCTIG

AGCACAAGAC

AGCTCCTGAG

5700 5760 5820 5880 5940 6000 6060 6120 6, Aq INFORMATION FOR SEQ ID NO:4: SEQUENCE CHARACTERISTICS: LENGTH: 926 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:

AAGGTAGGCA

AATATTAACA

C TrMATCAG

CAGTCCTI'AT

TAGTrAGT'r'

TCGGCTAACA

GAAGCTGGGA

GACAQGGTI'

TG-CCTCGGCC

CTA 'rTAAAT TTTGTCJ T

CTGAAGTCCG

TATCTCAGGA

ACGTGGTCTr

CCCJAAGGGGG

TTCTCTCT

CA'I~r.CCCTG CAATTTATAA AAAGTAGAGA GACTACAATG CACAGTCCAA TCATGTTTTA

CTAAAATCAA

TCAGhCASGAG

GCAATCTCCC

TrATAGACAC

CACCATTT

TCCCAAAGTG

GTCATCCTCA

TC-I TAATC

AGCTAACTGA

GTGGAGAGGG

TATCATGTC

AAArTAACI-r ATATCAAACA CCATACTCTr

GI'ICCCAAGC

CCTCCTTGvGT TCAAGCAATrr

TTTATCAGGT

AACTCCA.ATG

7 7ITCT1

CTGCCACCAC

GCCAGGCTGG

CTGGGATTAC

ACATAGTCAA

AGTTTCTACG

AGTTCCT13CT

AGACACCGGT

TAGTGCTGTG

CC!CTGGGAGC

CCTPGT

ATCC.A5CTAA TI'rCGAACTC

AGGCATGAGC

TCCTTGGGC

TGGAM TTGG

TCCCGAAT

ATGaAAG='AA 77CAGAGAAT

AGAGGGAGGG

GTTCAATTAT

TAGCCATAAC

TCTGhCCCCC

TGGGAGCCTA

TGGAGTACAA

CTCCTGCCTC

CTGACC TCAG

CACCACGCCT

A777rTTC= ACACTrTGGC

TGAAGGAGAG

GT77C'ITPCCC

CAGTATAGGG

GAGGA1GAAGA

GGAATTGTCA

TCAhGGCCCAA

AACCCATCCC

mM'1TTAGT

TAGTGTAGTC

AGTCTCCCAA

TTrAGAAAA

GTGATCCGCC

GGCCGGCADC

ACAGTAAAAT

CTTCCAGGAA

CCCATCTGGC

T1'rTGTGCCC

TAAATGCCCA

GGAACAGAAC

120 180 240 300 360 420 480 540 600 660 720 780 840 900 926 INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 2099 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: cDWM (xi) SEQUENCE DESCRIPTION: SEQ ID TGGCTCT1GCC AAtCCTrCcC CTCACGCTGA GAACAGCAG AAtCCCACCT ACCCCTACAC GTCCGCCAGG TITNTTGAGTA GTTCACATAC TGCCTAGQCC TATTTCCArGG GCGCTGAGTC AAGGCT GAGA AGGAAATCCC GCTACACGG AATTGACT GATGAGGCCA AAGGTOCCAT ATGATCAT~rx

TTTACCAIGA

AACAATTACT

CCAOCA"ACC

TGACCTCATC

ACTGGAAAGC

CAGAACTOTC

TGGAGCTGCC

GGCTGTGGXT

TGTCCTCTCC

TGTGGGAAC

GGCAAGTATG

ACGTGGAGAA

AGCCAGTTTA

ACGC4 TGCTG

ATAAGATATG

TACCACGGAC

GAAGCAGGCC

AAACT13AACC

GAAGATTATG

TAGTTTGGGT

GTUTUTGGAT

TCGACACA(GT

TGCAGGGCTA

TGGTQTACTC

AGCTGAATAC

AGTTCCCGTA

TCTGGGTCAT

CAGAGAATCbT GrATAGTGG AGGAaAGCCT

GCGAGACCCC

TOCACGGAT

CCCTTCTAAG

GGGGAGCCTC

CGAGACAGTG

j~wTCGCL,G

TTACAGCACC

GGAACTICGAA

120 180 240 300 360 420 480 540 600

CAAACCTGG

ACCTTYGTACA

ACAGGCACAG

AGCATGAMr GTCACrTATG

CAGAAGGAGA

GGCCCAGGCA

ACCATCTAAC

CCTTrATC7T 77'lGGGCAA

TACATGGTTA

TCTGGCCAGC

ATAGAATACA

L~-.7TV-ACLTCT

ATCAGGTAAT

CGCCAAGCAA

CCGGAGCAAT

AGT.LICTr

CACCCAGGCT

CACACCATTC

CCTGGCTAAT

TCGATCTCCT

GT13ACCCACC

TTATAAGCCA

AGACAAACAT

CCCTCAGCAG

GTATCAGCAA

ACTACAACCC

ACATCAAGCT

TGCTCAGGGC

GG~ACTG

TATTCAGGAA

CTGTCJAGCAT

AAGCTGTAAG

CCACAAGCCA

ATCGAATATA

CTTrAACTTT

TCCTTCAGGT

GTGTGTAACT

ATAGCGCATC

CTITrCCATCT

GGAGTGCAGT

TCCTGCCTCA

GACCTTGTCA

ACGCCCAGCC

CACCTCAGGT

CCGTAAGCAG

CTACACCTCA

GACCCTGACC

CCTGGAGAAG

CTCCAAGATG

TCCTGGGGGG

CTrr'CCAAGT

TTGTAGTCTG

TTATGGGATG

GCATAATAGT

CAATAAAAAG

AGTAAGATGC

ACATAACCCT

GTGrGGAATCT

TGGGAGATGT

CACACTTTCC

CTCCCCTGAA

GGCACGATCT

GCCTCCCAAG

TCCAGTGAAG

TCCACCCACC

CCT'CCACTI'C

GGAGAAAGCT

T CAGTCGCCA GCAGAT3CTA

ATCCCATTCA

AAGCTCTG

TGAAAAGCCT

AGCAGGCTGA

TTTCATrAAT

AGGGCGTAGA

TTIA.ATGACA

CTTTTCCT GA

CATAACTTCT

ATTACTACA

TACATP11TGA '171CTAAGA

GATIGCAGGA

TGC.C.TGGT7T~

CACCTCCATC

CCCACCCTCT

111TTTI

CGGCTCACTG

TAGCTGGGAC

ATGG'r=TA

TTGGCCTCCC

AGTTTTTATC

TGATGOCATAG

ATGCCTTCAT

CCGTCAACTT

AGAACCGCTA

CCTGGGACAA

CCAAGCTGTA

AGGGAGAGCC

CCAGAAGGAT

CAATI'TCATA

TAGTI'CAAGT

AAACCATTGC

AAAGGAAGCA

AATAAAATT

GAAGGATATA

TGTTAAAGGT

TCCTGT"GCT

ATTCACCCTA

GAGATGOCT

CAAGTTCCGC

TACAGGCACC

CCATTIAGC

AAAGTGCTGG

TGTCATCAGG

CTTGAGTATT

CATCGTGC

TGCTTATGAC

TAAGTACAGC

CTTAACATIG

CAGGCAATGG

AGCCAGCCAG

GAACATGGTC

TAATAAATAT

777CTGTA

TCTTGCATGT

GAATAGCTCC

AATGCTrrCAG

TCTTACCCAA

GA'rrCCAAcC

GSGJ'ZX

CCCAGGATGA

CTCGGCACTA

ACTCCACTTC

L 7 CZC.C GT CTCCCAGGTTr

TGCCACCACG

CAGGATGGTC

GATTACAGGC

GGTATV.AATT

CTATACTGT

660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2040 2099 INFORMATION FOR SEQ ID NO0:6: SEQUENCE CHARACTERISTICS: LENGTH: 19 base pairs TYPE: nucleic acid STRANDEDNESS: single TO POLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID 110:6: TGAGGCTTCC TCTGGAA.AC INFORMATION FOR SEQ ID 110:7: SEQUENCE CHARACTERISTICS: LENGTH: 20 bate pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID 110:7: TGAAATCAGC ACACCAGTAG INFORMATION FOR SEQ ID 110:8: SEQUENCE CHARCTERISTICS: LENGTH: 21 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8: GCACCCATAC CCCAATAATA G 21 INFORMATION FOR SEQ ID NO:9: SEQUENCE CHARACTERISTICS: LENGTH: 20 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: AGAGTTCCCC AGATTTCACC INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 20 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID ATCTGGGGAA CTCTTCTCAG INFORMATION FOR SEQ ID NO:11: SEQUENCE CHARACTERISTICS: LENGTH: 19 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11: TACAGTTGTT GCAGATACG 19 INFORMATION FOR SEQ ID NO:12: SEQUENCE CHARACTERISTICS: LENGTH: 21 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12: ACAACGTATC TGCAACAACT G 21 INFORMATION FOR SEQ ID NO:13: SEQUENCE CHARACTERISTICS: LENGTH: 20 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13: TCAGGCTTAA CTGCAGAACC "0 INFORMATION FOR SEQ ID NO:14: SEQUENCE CHARACTERISTICS: LENGTH: 19 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14: TTGGTTCTGC AGTTAAGCC 19 INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 19 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID AGCAGCACAA GGGCAATCC 19 INFORMATION FOR SEQ ID NO:16: SEQUENCE

CHARACTERISTICS:

LENGTH: 18 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16: ACAGGGCTAT ATTGTGGG 18 INFORMATION FOR SEQ ID NO:17: SEQUENCE CHARACTERISTICS: LENGTH: 19 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17: CCTGAGATGC CAGCTGTCC 19 INFORMATION FOR SEQ ID NO:18: SEQUENCE CHARACTERISTICS: LENGTH: 20 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18: CTGAAGCATT AGAAGCCAAC INFORMATION FOR SEQ ID NO:19: SEQUENCE CHARACTERISTICS: LENGTH: 20 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19: ACCTTGGACC AGGCTGCCAG INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 19 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID AGGTTTGTTC GAGTTCCAG 19 INFORMATION FOR SEQ ID NO:21: SEQUENCE CHARACTERISTICS: LENGTH: 20 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21: ACAATTACTG GCAAGTATGG INFORMATION FOR SEQ ID NO:22: SEQUENCE CHARACTERISTICS: LENGTH: 19 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22: CCT7VTCAGC C77GCTACC 19 INFORMATION FOR SEQ ID NO:23: nB r CATTC=!Jt- LENGTH: 20 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23: ACACCTCAGC AGATGCTACC INFORMATION FOR SEQ ID NO:24: SEQUENCE CHARACTERISTICS: LENGTH: 19 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24: ATGGATG3ACT GACATGGCC 19 S INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 19 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID AAGGATGAAC ATGGTCACC -19 INFORMATION FOR SEQ ID NO:26: Ci) SEQUENCE CHARACTE RISTICS: LENGTH: 1548 base pairs TYPE: nucleic acid STRANDEDNESS: single TOPOLOGY: .linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26: AGAGCTFTCC AGADGAAGCC TCACCAAGCC TCTGCAATGA GGTrCC1TCTG TGCACGTI'GC TGCAGC7N'G GGCCTGAGAT GCCAkGCTGTC CAGCTGCT0C TTCTGGCCTG CCTGGGTGMG 120 63

GATGTGGGGG

TATACCT11CA

TCAGTCATCC

AAAGCTCGAC

AGGCCCCAGG

GACCAGCTGG

AAGTCAGTTC

AGGTTGGAAA

CGAGACACTG

ACTTTGGCCT

AAGGAGAGC

GT'1Ir.GGTAG

GTGGGATGC

GACACAGTTG

CAGGGCTACC

GTGTACT-CGG

CTAATACCG

TTCCCGTATT

TGGGTCATT

GAGAATCTGG

GCCTrCATCA

GTCAACTT

AACCGCTATA

TGGGACAACT

CCAGGACAGC

GTGTG.GCCAG

ATAACTTACA

TCAGCTCCCT

AGACCCAGGA

AAACCCAAAC

TGGAGGAAGA

GCAGCAGCCA

ClZCGGCTGT~

TCCAGGAACT'

CAT='ZGCTA

GAGAGCCTCT

GAGACCCCAA

GCACGGATGT

CTTCTAAGGT

GGAGCCTCTA

AGACAGTGAA

ACAGCACCGA

AACTCGAACA

TCTGTGGCAC

CTI'ATGACAC

AGTACAGCAG

T3AACATGGT TCAGCTCAGG AAGGCCAT TrCCCAATGAA TCCAGCTGCC GACAGACAGC AGCACCCAAC GGAGAGCCTC CTCCACCAAT GGGGCTGCAG AGGGAGCTGG CAGAGAGTTG GAGACTGCCT GAAGAAGCGA CTAAGGCAAG GGAGGTAGCA AGGCTGAGAA GCCACCAGGC TCCAGAGAAG TC'rCAGGAGT GGAGAGGGAG CACGCTGAGA ACAGCAGAAA GCCCACCTAC CCCTACACCC CCGCCAGGTT TTTGAGTATG TCACATACTG CCTAGGCCAC T1-CCAGGGC GCTGAGTCCA GGCTGAGAAG GAAATCCCTG CTACACGGAC ATTGACTG TGAGGCCAAA GGTGCCATTG AACC!TGGGAG ACAAACATCC CTTGTACACC GTCAGCAGCT AGGCACAGCT ATCAGCAAGA CATGATTGAC TACAACCCCC CACITATGAC ATCAAGCTCT

ACCAGAGTGG

CAaAGCAGAG

GCTTAGACCT

TGACCTTGGA

GCAcCCTGAG

ACAGCAACCT

AAAATGAGAA

GGGGCCAGT

TP'rCTACGTG

ACACCGGATG

CA7LTrACTGG

AGGAGACCAC

ACCTCATCAG

TGGAAAGCAC

GAACTGTCAT

GAGCTGGCTA

CTGTGATGA

TcCC~CAA

GTAAGCAGTC

ACACCTC.AGC!

CCCTGACCAT

TGGAGAA GAA

CCAAGATG

CCGATGCCAG

ccAGGCCATG

GGAGGCCACC

CCAGGCTGcc

GCGGGAGCGG

CCTICCGAGAC

TCTGGCCAGG

TiCCCCAGACC

GAATTTGGAC

CCGAkI-jGc

'GGAGAACTA

CAAGTATIGGT

GTGGAGAATC

CCAG~rTATG

GGGTGCTG

AAGATATGAG

CCACGGACAG

AGCAGGCCTc AcTGAAcccA

AGTCGCCAAT

AGATGCTACC

CCCATTCAAG

GCTCTT'rGCC 180 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1548

S

S. S S S

S.

*5*S INFORMATION FOR SEQ ID NO:27: SEQUENCE CHARACTERISTICS: LENGTH: 178 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: None (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27: Thr 1 Ser Gly Ala Val Val Tyr Ser Arg Thr Val Ile Arg Tyr Pro Gly Ala Gly Ser Leu 10 Giu Lou Asn 25 Gly Tyr His Tyr Phe Gin Gly Ala Glu is Thr Glu Thr Glu Lys Giu Trp Gly Gly Ile Gly Gin Val Lys Ala Pro Tyr Ser Ala Gly Lou Tyr Thr Asp *5 55 S so Trp Val Ile Asp Lou Ala Val le Tyr Ser Lys Thr Asp 70 Asn Leu Glu Ala Lys Gly 75 Ile Val Lou Ser so Lou Asn Pro Giu Ser Glu Lou Glu Gin Thr Trp Glu Thr Asn Ile Arg Lys Gin 100 Tyr Thr Val Ser 115 Tyr Asp Thr Gly Val Ala Asn Phe Ile Ile Cys Ser Tyr Thr Ser 120 Ala Asp Ala Thr Val 125 Ile Gly Thr Lou 110 Asn Phe Ala Pro Phe Lys Thr Gi 130 Asn Arg y Ile Ser .135 r Ser Met Tyr Lys Tyr Se 145 Lys 150 Asp Lys Thr Lou Ile Asp Tyr 155 Asn Met Val 170 Thr 140 Asn Pro Lou Giu Lys 160 Lys Lou Phe Ala Trp 165 Asn Lou Thr Tyr Asp Lou Ser INFORMATION FOR SEQ ID NO:28: SEQUENCE CHARACTERISTICS: LENGTH: 131 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: None Arg 1 Tyr Asp Thr Ser Pro Tyr Thr Phe (xi) SEQUENCE Phe Asp Leu Lys 5 Ala Gly Tyr Asn Ile Asp Leu Met Asn Gin Asn Ala Leu Gin Thr Leu Gly Xaa Ala Phe 85 Ser Gly Gly Thr 100 Tyr Glu Tyr Ile 115 Pro Cys 130

DESCRIPTION:

Thr Glu Thr Ile Leu Lys Thr Arg Ser Leu SEQ ID NO:28: Tyr Glu 40 Ile Trp Cys His Pro 120 Trp Trp Arg Tyr Tyr Gin Lys Gly Ala Leu Pro Val Thr Leu 125 Gly Val Asp Lys Thr Asn 110 Xaa His Tyr Pro Arg Asn Ala Pro Asp Ser Ala Val Xaa Gly Ser His 0 eeoc

S

0

S

0 00 She.

INFORMATION FOR SEQ ID NO:29: SEQUENCE CHARACTERISTICS: LENGTH: 178 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: None 0*

S

Gly 1 Gin Lys Ala Leu Ser Ser Leu (xi) SEQUENCE Thr Gly Gin Val 5 Ser His Ile Ile Thr Arg Ser Leu Trp Gly Gly His Trp Ala Val Tyr Lys Leu Asp Pro Tyr Pro Lys Arg 100 Tyr Val Thr Asn DESCRIPTION: SEQ ID Val Tyr Asn Gly Ser Ile Arg Phe Asp Leu Asp Tyr Ala Gly Tyr Ser Asp Ile Asp Leu 55 Ala Thr Asn Gin Asn 70 Val Ser Leu Gin Ile Ser Ala Gly Glu Ala 105 Gly Tyr Ser Gly Gly NO:29: Ile Tyr Lys Thr Asn Asn Met Val Ala Gly Leu Gin Phe Ile Thr Lys 115 120 125 Tyr Gin Thr Asn Ala Ser Thr Tyr Glu Tyr Ile Asp Ile Pro Phe Gin 130 135 140 Asn Lys Tyr Ser His Ile Ser Met Leu Asp Tyr Asn Pro Lys Asp Arg 145 150 155 160 Ala Leu Tyr Ala Trp Asn Asn Gly His Gin Thr Leu Tyr Asn Val Thr 165 170 175 Leu Phe INFORMATION FOR SEQ ID SEQUENCE CHARACTERISTICS: LENGTH: 177 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: None (xi) SEQUENCE DESCRIPTION: SEQ ID Gly Ala Gly Val Val Val His Asn Asn Asn Leu Tyr Tyr Asn Cys Phe 1 5 10 S Asn Ser His Asp Met Cys Arg Ala Ser Leu Thr Ser Gly Val Tyr Gin 25 Lys Lys Pro Leu Leu Asn Ala Leu Phe Asn Asn Arg Phe Ser Tyr Ala 35 40 Gly Thr Met Phe Gin Asp Met Asp Phe Ser Ser Asp Glu Lys Gly Leu 55 Trp Val Ile Phe Thr Thr Glu Lys Ser Ala Gly Lys Ile Val Val Gly 70 75 Lys Val Asn Val Ala Thr Phe Thr Val Asp Asn Ile Trp Ile Thr Thr 90 Gin Asn Lys Ser Asp Ala Ser Asn Ala Phe Met Ile Cys Gly Val Leu 100 105 110 Tyr Val Thr Arg Ser Leu Gly Pro Lys Met Glu Glu Val Phe Tyr Met 115 120 125 Phe Asp Thr Lys Thr Gly Lys Glu Gly His Leu Ser Ile Met Met Glu 130 135 140 Lys Met Ala Glu Lys Val His Ser Leu Ser Tyr Asn Ser Asn Asp Arg 145 150 155 160 Lys Leu Tyr Met Phe Ser Glu'Gly Tyr Leu Leu His Tyr Asp Ile Ala 165 170 175 Leu INFORMATION FOR SEQ ID NO:31: SEQUENCE CHARACTERISTICS: LENGTH: 74 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: None (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31: Gly Val Val Tyr Ser Arg Leu Thr Glu Thr Leu Ala Gly Tyr Asn Asn 66 1 Tyr Ala Thr Al a Phe Ile Ile Tyr Trp Gly Ile Asp

S

Gly Gly Asp Ile Asp Lou 25 Ile Val Ser Lys Leu Pro 40 Cys Gly Thr Lou Tyr Val 55 Tyr Asn Pro Lys Lou Tyr 10 Val Asp Glu Gly Lou Ti-p Tyr Leu Gin Th- Ti-p Thr Lys Ala Thr Tyr Val Tyr Ala Tyr Thr Lou INFORMATION FOR SEQ ID NO:32: SEQUENCE CHARACTERISTICS: LENGTH: 504 amino acids TYPE: amino acid STRANDEDNESS: single TOPOLOGY: linear (ii) MOLECULE TYPE: protein FRAGMENT TYPE: N-terminal (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32: Met 1 Ala Arg Tyr Ser Gin Ser Th- Asp Lou 145 Gin Val Arg Thr Ser 225 Gly Lou Asp Arg Val Th- Th- 50 Gin Arg Lou Gin Gin- 130 Lou Giu Ala Ala Lou 210 Arg Asp Arg Pro Phe Phe Cys Ala 5 Gin Leu Leu Lou 20 Ala Gin Leu Arg 40 Phe Ser Val Ala Ala Met Ser Val 70 Lou Asp Lou Glu 85 Lou His Gin Lou 100 Glu Giy Lou Gin 115 Lou Giu Thr Gin Arg Asp Lys Ser 150 Asn Glu Asn Lou 165 Arg Lou Arg Arg 180 Val Pro Pro Gly 195 Ala Phe Gin Glu Ile Lou Lys Glu 230 Thr Gly Cys Gly 245 Thr Ala Glu Thr 260 Lys Pro Thr Tyr 275 Arg Lou Lys Ser 55 Ile Ala Th- Arg Thr 135 Val Ala Giy Ser Lou 215 Ser Giu Ile Pro Cys Ala Ala Pro His Thr Lou Giu 120 Arg Lou Arg Gin Arg 200 Lys Pro Lou Th- Tyr 280 Cys Cys 25 Asn Asn Asn Lys Asp 105 Lou Glu Glu Arg Cys 185 Glu Ser Ser Val Gly 265 Th- Ser Phe Lou Val Asp Gin Giu Ser Lou Gin 75 Ala Arg 90 Gin Ala Gly Thr Lou Glu Glu Giu 155 Lou Giu 170 Pro Gin Val Ser Glu Lou Gly Tyr 235 Ti-p Val 250 Lys Tyr Gin Glu Val Phe 67 Gly Ti-p Ser Ser Arg Lou Ala Leu Th- 140 Lys Ser Thr Th- Thr 220 Leu Gly Gly Th- Glu Pro Glu Asp Val Gly Arg Cys Pro Asp Ser Ser Ser Arg Pro 110 Arg Arg 125 Ala Tyr Lys Arg Ser Ser Arg Asp 190 Ti-p Asn 205 Glu Val Arg Ser Giu Pro Val Trp 270 Thr Ti-p 285 Tyr Asp Met Gly Cys Giu Ser Lou Gin Giu Ser Lou Gin 175 Thr Lou Pro Gly Lou 255 Met Arg Leu Pro Ala Gin Gin Th- Glu Glu Arg Asn Arg 160 Glu Ala Asp Ala Giu 240 Thr Arg Ile Ile Asp Thr Val Gly Th- Asp Val Arg Gin 290 295 300 Ser Gin Phe Met Gin Gly Tyr Pro Ser Lys Val His Ile Leu Pro Arg 305 310 315 320 Pro Leu Glu Ser Thr Gly Ala Val Val Tyr Ser Gly Ser Leu Tyr Phe 325 330 335 Gin Gly Ala Giu Ser Arg Thr Val Ile Arg Tyr Giu Leu Asn Thr Giu 340 345 350 Thr Val Lys Ala Giu Lys Glu Ile Pro Gly Ala Gly Tyr His Gly Gin 355 360 365 Phe Pro lyr Ser Trp, Gly Gly Tyr Thr Asp Ile Asp Leu Ala Val Asp 370 375 380 Giu Ala Gly Leu 'rrp Vai Ile Tyr Ser Thr Asp Giu Ala Lys Gly Ala 385 390 395 400 Ile Val Leu Ser Lys Leu Asn Pro Giu Asn Leu Giu Leu Giu Gin Thr 405 410 415 Trp Giu Thr Asn Ile Arg Lys Gin Ser Vai Ala Asn Ala Phe le Ile 420 425 430 Cys Gly Thr Leu Tyr Thr Val Ser Ser Tyr Thr Ser Ala Asp Ala Thr 435 440 445 Val Asn Phe Ala Tyr Asp Thr Gly Thr Gly Ile Ser Lys Thr Leu Thr 450 455 460 Ile Pro Phe Lys Asn Arg Tyr Lys Tyr Ser Ser Met Ile Asp Tyr Asn 465 470 475 480 Pro Leu Giu Lys Lys Leu Phe Ala Trp Asp Asn Leu Asn Met Val Thr 485 490 495 Tyr Asp Ile Lys Leu Ser Lys Met 500

Claims

1. A composition which comprises: a first marker nucleic acid molecule, said first marker nucleic acid molecule comprising a nucleotide sequence of a polynucleotide that specifically hybridizes to a polynucleotide comprising the sequence of SEQ ID NO: 2 or its complement; and a complementary nucleic acid molecule obtained from a cell or a bodily fluid of a patient, wherein nucleic acid hybridization between said first marker nucleic acid molecule and said complementary nucleic acid molecule obtained from said patient permits the detection of a polymorphism whose presence is predictive of a mutation affecting TIGR response in said patient.

2. The composition according to claim 1, wherein said first marker nucleic acid molecule is capable of specifically detecting TIGRmtl.

3. The composition according to claim 1, wherein said first marker nucleic acid molecule is capable of specifically detecting TIGRmt2. S: 20

4. The composition according to claim 1, wherein said first marker nucleic acid molecule is capable of specifically detecting TIGRmt3. o :o:

5. The composition according to claim 1, wherein said first marker nucleic acid molecule is capable of specifically detecting TIGRmt4.

6. The composition according to claim 1, wherein said first marker nucleic acid molecule is capable of specifically detecting TIGRmtS.

7. The composition according to claim 1, wherein said first marker nucleic acid molecule is capable of specifically detecting TIGRsvl.

8. The composition according to claim 1, further comprising a second marker nucleic acid molecule.

9. The composition according to claim 8, wherein said first marker nucleic acid molecule and said second marker nucleic acid molecule are selected from the group consisting of a nucleic acid molecule that comprises a sequence selected from the group consisting of: SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO:

10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16 and their complements. The composition according to claim 9, wherein said first marker nucleic acid molecule and said second marker nucleic acid molecule are selected from the group consisting of a nucleic acid molecule that comprises a sequence selected from the group consisting of: SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, and their complements.

11. The composition according to any of claims 1 through 10, wherein said complementary nucleic acid molecule has been subject to nucleotide amplification.

12. The composition according to claim 11, wherein said nucleotide amplification is conducted by means of polymerase chain reaction.

•13. The composition according to claim 11, wherein said nucleotide amplification is conducted by means of oligonucleotide ligation assay.

14. The composition according to claim 11, capable of detecting single nucleotide polymorphisms in said complementary nucleic acid. The composition according to any one of claims 1 to 14, substantially as hereinbefore described with reference to any of the examples and/or drawings. Dated this seventh day of December 2004 The Regents of the University of California Patent Attorneys for the Applicant: F B RICE CO