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WO2009050498A3 - Chimeric constructs for eukaryotic membrane expression in prokaryotic cells - Google Patents

Chimeric constructs for eukaryotic membrane expression in prokaryotic cells Download PDF

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Publication number
WO2009050498A3
WO2009050498A3 PCT/GB2008/003573 GB2008003573W WO2009050498A3 WO 2009050498 A3 WO2009050498 A3 WO 2009050498A3 GB 2008003573 W GB2008003573 W GB 2008003573W WO 2009050498 A3 WO2009050498 A3 WO 2009050498A3
Authority
WO
WIPO (PCT)
Prior art keywords
membrane protein
prokaryotic cells
chimeric constructs
membrane expression
chimeric membrane
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/GB2008/003573
Other languages
French (fr)
Other versions
WO2009050498A2 (en
Inventor
Bonnie Ann Wallace
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
BIRKBECK
Original Assignee
BIRKBECK
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 BIRKBECK filed Critical BIRKBECK
Publication of WO2009050498A2 publication Critical patent/WO2009050498A2/en
Publication of WO2009050498A3 publication Critical patent/WO2009050498A3/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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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
    • C12N15/63—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
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/62—DNA sequences coding for fusion proteins
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/03—Fusion polypeptide containing a localisation/targetting motif containing a transmembrane segment

Landscapes

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

Abstract

An isolated nucleic acid molecule encoding a chimeric membrane protein is provided. The chimeric membrane protein comprises at least two N-terminal transmembrane segments of a prokaryotic membrane protein and the remaining transmembrane segments are eukaryotic. A chimeric membrane protein is also provided. The nucleic acid molecule encoding the chimeric membrane protein can be inserted into a prokaryote, for example, a bacterium such as E. coli. This can be done by introducing a vector into the prokaryote. The chimeric membrane protein encoded by the nucleic acid molecule is then expressed in the prokaryote.
PCT/GB2008/003573 2007-10-19 2008-10-20 Chimeric constructs for eukaryotic membrane expression in prokaryotic cells Ceased WO2009050498A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB0720563.6 2007-10-19
GB0720563A GB0720563D0 (en) 2007-10-19 2007-10-19 Protein

Publications (2)

Publication Number Publication Date
WO2009050498A2 WO2009050498A2 (en) 2009-04-23
WO2009050498A3 true WO2009050498A3 (en) 2009-06-04

Family

ID=38814197

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB2008/003573 Ceased WO2009050498A2 (en) 2007-10-19 2008-10-20 Chimeric constructs for eukaryotic membrane expression in prokaryotic cells

Country Status (2)

Country Link
GB (1) GB0720563D0 (en)
WO (1) WO2009050498A2 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB0823366D0 (en) * 2008-12-22 2009-01-28 Uni I Oslo Synthesis

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003040323A2 (en) * 2001-11-08 2003-05-15 Children's Medical Center Corporation Bacterial ion channel and a method for screening ion channel modulators
US20050202539A1 (en) * 2004-03-11 2005-09-15 Hydra Biosciences System for screening eukaryotic membrane proteins

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003040323A2 (en) * 2001-11-08 2003-05-15 Children's Medical Center Corporation Bacterial ion channel and a method for screening ion channel modulators
US20050202539A1 (en) * 2004-03-11 2005-09-15 Hydra Biosciences System for screening eukaryotic membrane proteins

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
DATABASE Geneseq [online] 2 September 2003 (2003-09-02), "Bacterial voltage-sensitive ion-selective channel protein NaChBac.", XP002516535, retrieved from EBI accession no. GSP:ABR55917 Database accession no. ABR55917 *
FU D ET AL: "Structure of a glycerol-conducting channel and the basis for its selectivity", SCIENCE 2000 US, vol. 290, no. 5491, 2000, pages 481 - 486, ISSN: 0036-8075 *
IRENE NEOPHYTOU ET AL: "Eukaryotic integral membrane protein expression utilizing the Escherichia coli glycerol-conducting channel protein (GlpF)", APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, SPRINGER, BERLIN, DE, vol. 77, no. 2, 9 September 2007 (2007-09-09), pages 375 - 381, XP019560704, ISSN: 1432-0614 *
LAAGE R ET AL: "Strategies for prokaryotic expression of eukaryotic membrane proteins", TRAFFIC, MUNKSGAARD, DK, vol. 2, no. 2, 1 February 2001 (2001-02-01), pages 99 - 104, XP009023195, ISSN: 1398-9219 *
LU ZHE ET AL: "Ion conduction pore is conserved among potassium channels", NATURE, vol. 413, 25 October 2001 (2001-10-25), pages 809 - 813, XP002517880, ISSN: 0006-3495 *

Also Published As

Publication number Publication date
GB0720563D0 (en) 2007-11-28
WO2009050498A2 (en) 2009-04-23

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