WO1996009378A1 - Overexpression of mammalian and viral proteins - Google Patents
Overexpression of mammalian and viral proteins Download PDFInfo
- Publication number
- WO1996009378A1 WO1996009378A1 PCT/US1995/011511 US9511511W WO9609378A1 WO 1996009378 A1 WO1996009378 A1 WO 1996009378A1 US 9511511 W US9511511 W US 9511511W WO 9609378 A1 WO9609378 A1 WO 9609378A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- gene
- protein
- seq
- codons
- synthetic
- Prior art date
Links
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
- C07K14/43595—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from coelenteratae, e.g. medusae
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/70503—Immunoglobulin superfamily
-
- 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
- C12N15/67—General methods for enhancing the expression
-
- 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
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
-
- 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
- C12N2740/00—Reverse transcribing RNA viruses
- C12N2740/00011—Details
- C12N2740/10011—Retroviridae
- C12N2740/16011—Human Immunodeficiency Virus, HIV
- C12N2740/16111—Human Immunodeficiency Virus, HIV concerning HIV env
- C12N2740/16122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
Definitions
- Figure 4 is a graph depicting the results of ELISA assays used to measure protein levels in supernatants of transiently transfected 293T cells.
- cytoplasmic mRNA 293 cells transfected with CDM&, gpl20 IIIB, or syngpl20 was isolated 36 hours post transfection.
- Cytoplasmic RNA of Hela cells infected with wildtype vaccinia virus or recombinant virus expressing gpl20 Illb or the synthetic gpl20 gene was under the control of the 7.5 promoter was isolated 16 hours post infection.
- Equal amounts were spotted on nitrocellulose using a slot blot device and hybridized with randomly labelled 1.5 kb gpl20IIIb and syngpl20 fragments or human beta-actin. RNA expression levels were quantitated by scanning the hybridized membranes with a phospoimager. The procedures used are described in greater detail below.
- the expected number of CpG dinucleotides for HIV as a whole is about one fifth that expected on the basis of the base composition. This might indicate that the possibility of high expression is restored, and that the gene in fact has to be highly expressed at some point during viral pathogenesis.
- the results presented herein clearly indicate that codon preference has a severe effect on protein levels, and suggest that translational elongation is controlling mammalian gene expression. However, other factors may play ar role.
- abundance of not maximally loaded mRNA's in eukaryotic cells indicates that initiation is rate limiting for translation in at least some cases, since otherwise all transcripts would be completely covered by ribosomes.
- Transformed bacteria were grown in 3 ml LB cultures for more than 6 hours or overnight.
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Chemistry (AREA)
- Immunology (AREA)
- Toxicology (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Peptides Or Proteins (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Compounds Of Unknown Constitution (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
Claims
Priority Applications (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP95931798A EP0781329B2 (en) | 1994-09-19 | 1995-09-08 | Overexpression of mammalian and viral proteins |
DE69535147T DE69535147T3 (en) | 1994-09-19 | 1995-09-08 | OVEREXPRESSION OF MAMMAL AND VIRUS PROTEINS |
CA002200342A CA2200342C (en) | 1994-09-19 | 1995-09-08 | Overexpression of mammalian and viral proteins |
JP8510951A JPH10506278A (en) | 1994-09-19 | 1995-09-08 | Overexpression of mammalian and viral proteins |
AU35099/95A AU3509995A (en) | 1994-09-19 | 1995-09-08 | Overexpression of mammalian and viral proteins |
DK95931798.3T DK0781329T4 (en) | 1994-09-19 | 1995-09-08 | Overexpression of mammalian and viral proteins |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US08/324,243 | 1994-09-19 | ||
US08324243 US5786464C1 (en) | 1994-09-19 | 1994-09-19 | Overexpression of mammalian and viral proteins |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1996009378A1 true WO1996009378A1 (en) | 1996-03-28 |
Family
ID=23262728
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US1995/011511 WO1996009378A1 (en) | 1994-09-19 | 1995-09-08 | Overexpression of mammalian and viral proteins |
Country Status (13)
Country | Link |
---|---|
US (1) | US5786464C1 (en) |
EP (1) | EP0781329B2 (en) |
JP (1) | JPH10506278A (en) |
AT (1) | ATE335078T1 (en) |
AU (1) | AU3509995A (en) |
CA (1) | CA2200342C (en) |
DE (1) | DE69535147T3 (en) |
DK (1) | DK0781329T4 (en) |
ES (1) | ES2270430T5 (en) |
PT (1) | PT781329E (en) |
TR (1) | TR199501134A2 (en) |
WO (1) | WO1996009378A1 (en) |
ZA (1) | ZA957846B (en) |
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WO1998012207A1 (en) * | 1996-09-20 | 1998-03-26 | The General Hospital Corporation | High level expression of proteins |
EP0851868A1 (en) * | 1995-09-22 | 1998-07-08 | The General Hospital Corporation | High level expression of proteins |
WO1998034640A2 (en) * | 1997-02-07 | 1998-08-13 | Merck & Co., Inc. | Synthetic hiv gag genes |
EP1002091A1 (en) * | 1997-07-09 | 2000-05-24 | The University Of Queensland | Nucleic acid sequence and method for selectively expressing a protein in a target cell or tissue |
WO2000039304A2 (en) * | 1998-12-31 | 2000-07-06 | Chiron Corporation | Polynucleotides encoding antigenic hiv type c polypeptides, polypeptides and uses thereof |
WO2001068835A2 (en) * | 2000-03-13 | 2001-09-20 | Aptagen | Method for modifying a nucleic acid |
WO2002008417A1 (en) * | 2000-07-25 | 2002-01-31 | Takeda Chemical Industries, Ltd. | Process for producing recombinant protein |
JP2002503477A (en) * | 1998-02-17 | 2002-02-05 | オックスフォード バイオメディカ(ユーケイ)リミテッド | Antiviral vector |
WO2002042326A1 (en) * | 2000-11-22 | 2002-05-30 | Biota Scientific Management Pty Ltd | A method of expression and agents identified thereby |
WO2002083181A1 (en) | 2001-04-18 | 2002-10-24 | The University Of Queensland | Novel compositions and uses therefor |
US6489141B1 (en) * | 1997-07-09 | 2002-12-03 | The University Of Queensland | Nucleic acid sequence and methods for selectively expressing a protein in a target cell or tissue |
US6534312B1 (en) | 1996-02-22 | 2003-03-18 | Merck & Co., Inc. | Vaccines comprising synthetic genes |
WO2002064799A3 (en) * | 1999-09-28 | 2003-03-27 | Transkaryotic Therapies Inc | Optimized messenger rna |
WO2003025003A2 (en) * | 2001-09-20 | 2003-03-27 | Glaxo Group Limited | Hiv-gag codon-optimised dna vaccines |
US6649409B1 (en) | 1999-03-29 | 2003-11-18 | Statens Serum Institut | Method for producing a nucleotide sequence construct with optimized codons for an HIV genetic vaccine based on a primary, early HIV isolate and synthetic envelope BX08 constructs |
US6656706B2 (en) | 1999-12-23 | 2003-12-02 | The United States Of America As Represented By The Department Of Health And Human Services | Molecular clones with mutated HIV gag/pol, SIV gag and SIV env genes |
US6696291B2 (en) | 1997-02-07 | 2004-02-24 | Merck & Co., Inc. | Synthetic HIV gag genes |
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US6924365B1 (en) | 1998-09-29 | 2005-08-02 | Transkaryotic Therapies, Inc. | Optimized messenger RNA |
WO2008049782A1 (en) * | 2006-10-24 | 2008-05-02 | Basf Se | Method of increasing gene expression using modified codon usage |
US7935805B1 (en) | 1998-12-31 | 2011-05-03 | Novartis Vaccines & Diagnostics, Inc | Polynucleotides encoding antigenic HIV Type C polypeptides, polypeptides and uses thereof |
US8133494B2 (en) | 2001-07-05 | 2012-03-13 | Novartis Vaccine & Diagnostics Inc | Expression cassettes endcoding HIV-1 south african subtype C modified ENV proteins with deletions in V1 and V2 |
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Also Published As
Publication number | Publication date |
---|---|
DK0781329T3 (en) | 2006-12-11 |
CA2200342C (en) | 2009-03-03 |
ES2270430T3 (en) | 2007-04-01 |
EP0781329B2 (en) | 2010-12-29 |
ATE335078T1 (en) | 2006-08-15 |
DE69535147T3 (en) | 2011-07-07 |
EP0781329A1 (en) | 1997-07-02 |
AU3509995A (en) | 1996-04-09 |
PT781329E (en) | 2006-12-29 |
EP0781329A4 (en) | 1998-11-25 |
ES2270430T5 (en) | 2011-05-12 |
EP0781329B1 (en) | 2006-08-02 |
DE69535147D1 (en) | 2006-09-14 |
DE69535147T2 (en) | 2007-07-05 |
US5786464A (en) | 1998-07-28 |
CA2200342A1 (en) | 1996-03-28 |
DK0781329T4 (en) | 2011-01-24 |
ZA957846B (en) | 1997-03-18 |
JPH10506278A (en) | 1998-06-23 |
US5786464C1 (en) | 2012-04-24 |
TR199501134A2 (en) | 1996-06-21 |
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