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WO2003004599A3 - Methods of making and using expression constructs for the expression of recombinant proteins - Google Patents

Methods of making and using expression constructs for the expression of recombinant proteins Download PDF

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Publication number
WO2003004599A3
WO2003004599A3 PCT/IL2002/000540 IL0200540W WO03004599A3 WO 2003004599 A3 WO2003004599 A3 WO 2003004599A3 IL 0200540 W IL0200540 W IL 0200540W WO 03004599 A3 WO03004599 A3 WO 03004599A3
Authority
WO
WIPO (PCT)
Prior art keywords
expression
fusion polypeptide
making
methods
recombinant proteins
Prior art date
Application number
PCT/IL2002/000540
Other languages
French (fr)
Other versions
WO2003004599A2 (en
Inventor
Yoav Peleg
Zeev Pancer
Original Assignee
Insight Biopharmaceuticals Ltd
Yoav Peleg
Zeev Pancer
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Insight Biopharmaceuticals Ltd, Yoav Peleg, Zeev Pancer filed Critical Insight Biopharmaceuticals Ltd
Priority to AU2002311620A priority Critical patent/AU2002311620A1/en
Publication of WO2003004599A2 publication Critical patent/WO2003004599A2/en
Publication of WO2003004599A3 publication Critical patent/WO2003004599A3/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/62—DNA sequences coding for fusion proteins
    • C12N15/625—DNA sequences coding for fusion proteins containing a sequence coding for a signal sequence
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/74—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/034—Fusion polypeptide containing a localisation/targetting motif containing a motif for targeting to the periplasmic space of Gram negative bacteria as a soluble protein, i.e. signal sequence should be cleaved
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/10—Fusion polypeptide containing a localisation/targetting motif containing a tag for extracellular membrane crossing, e.g. TAT or VP22
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
    • C07K2319/74—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor
    • C07K2319/75—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor containing a fusion for activation of a cell surface receptor, e.g. thrombopoeitin, NPY and other peptide hormones

Landscapes

  • Genetics & Genomics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

A method of producing a fusion polypeptide in, and purifying the fusion polypeptide from, bacteria, the method comprises introducing into the bacteria an expression construct encoding the fusion polypeptide which comprises a TAT-derived peptide, and a protein-of-interest, whereby the TAT-derived peptide serves for transport of the fusion polypeptide from the bacterial cytoplasm to the periplasm. The fusion polypeptide is then substantially exclusively purified from the bacterial periplasm.
PCT/IL2002/000540 2001-07-02 2002-07-02 Methods of making and using expression constructs for the expression of recombinant proteins WO2003004599A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2002311620A AU2002311620A1 (en) 2001-07-02 2002-07-02 Methods of making and using expression constructs for the expression of recombinant proteins

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US30180501P 2001-07-02 2001-07-02
US60/301,805 2001-07-02

Publications (2)

Publication Number Publication Date
WO2003004599A2 WO2003004599A2 (en) 2003-01-16
WO2003004599A3 true WO2003004599A3 (en) 2004-03-18

Family

ID=23164962

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IL2002/000540 WO2003004599A2 (en) 2001-07-02 2002-07-02 Methods of making and using expression constructs for the expression of recombinant proteins

Country Status (2)

Country Link
AU (1) AU2002311620A1 (en)
WO (1) WO2003004599A2 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102766645B (en) * 2012-06-08 2013-06-05 武汉凯肽来生物科技有限公司 Oral recombinant protein TAT-GH for promoting ricefield eel growth, and preparation method and application thereof

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2391457T3 (en) * 2003-08-13 2012-11-26 Sandoz Ag Procedure for purification of recombinant polypeptides
ES2281822T3 (en) 2003-08-13 2007-10-01 Sandoz Ag EXPRESSION VECTORS, TRANSFORMED GUEST CELLS AND FERMENTATION PROCEDURE FOR THE PRODUCTION OF RECOMBINATING POLYPEPTIDES.
GB0319601D0 (en) * 2003-08-20 2003-09-24 Sandoz Ag Production process
WO2014084432A1 (en) * 2012-11-30 2014-06-05 경상대학교 산학협력단 Helicobacter pylori expression vector
CN111705032B (en) * 2020-07-09 2022-04-05 科赫生物科技（北京）有限公司 Prokaryotic expression cell and preparation method and application thereof
CN118562843B (en) * 2024-08-05 2024-11-19 南京永正生物科技有限公司 Preparation method and application of a high-stability fusion enzyme

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6221355B1 (en) * 1997-12-10 2001-04-24 Washington University Anti-pathogen system and methods of use thereof
US6485977B1 (en) * 1999-09-13 2002-11-26 Cornell Research Foundation, Inc. Recombinant constructs and techniques for delivering to eucaryotic cells bacterial proteins that are secreted via type III secretion systems
US20030018001A1 (en) * 1996-11-21 2003-01-23 Heaney Anthony P. Methods of using pituitary tumor transforming gene (PTTG) carboxy-terminal peptides to inhibit neoplastic cellular proliferation and/or transformation of breast and ovarian cells

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030018001A1 (en) * 1996-11-21 2003-01-23 Heaney Anthony P. Methods of using pituitary tumor transforming gene (PTTG) carboxy-terminal peptides to inhibit neoplastic cellular proliferation and/or transformation of breast and ovarian cells
US6221355B1 (en) * 1997-12-10 2001-04-24 Washington University Anti-pathogen system and methods of use thereof
US6485977B1 (en) * 1999-09-13 2002-11-26 Cornell Research Foundation, Inc. Recombinant constructs and techniques for delivering to eucaryotic cells bacterial proteins that are secreted via type III secretion systems

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102766645B (en) * 2012-06-08 2013-06-05 武汉凯肽来生物科技有限公司 Oral recombinant protein TAT-GH for promoting ricefield eel growth, and preparation method and application thereof

Also Published As

Publication number Publication date
WO2003004599A2 (en) 2003-01-16
AU2002311620A1 (en) 2003-01-21

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