Process for production of polypeptides
AssigneeGenentech, Inc.
InventorsEriks Sasha Paegle, Dorothea Reilly, Daniel G. Yansura
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Abstract
Vectors for producing polypeptides heterologous to prokaryotes are described comprising, along with the polypeptide-encoding nucleic acid, anti-termination nucleic acid that inhibits intragenic transcription termination with a non-lambda promoter therefor and/or nucleic acid encoding a GreA or GreB protein and a promoter therefor. Also described are processes for producing a heterologous polypeptide in prokaryotic host cells utilizing such elements to improve the quality and/or quantity of heterologous polypeptide produced.
Claims30
A vector for producing a polypeptide heterologous to prokaryotic cells comprising (1) anti-termination nucleic acid that inhibits intragenic transcription termination with a non-lambda promoter therefor, (2) DNA encoding the polypeptide with a non-lambda promoter therefor, wherein an RNA recognition site for binding anti-termination protein produced from the nucleic acid is located 5′ of the DNA encoding the polypeptide, and (3) nucleic acid encoding a GreA or GreB protein with a promoter therefor.
A process for increasing production of a full-length heterologous polypeptide as a percentage of total such heterologous polypeptide in prokaryotic host cells comprising:
- (a) culturing the host cells, which comprise (1) anti-termination nucleic acid that inhibits intragenic transcription termination with a non-lambda promoter therefor, and (2) RNA encoding the polypeptide wherein the RNA is expressed from a gene with a non-lambda promoter therefor, wherein an RNA recognition site for binding anti-termination protein produced from the nucleic acid is located 5′ of the RNA encoding the polypeptide, and wherein the anti-termination nucleic acid is expressed at the time of expression of the RNA;
- and
- (b) recovering the heterologous polypeptide from the cells or from cell culture medium, whereby the amount of full-length heterologous polypeptide produced by the process is increased as a percentage of total said heterologous polypeptide produced.
A process for increasing production of a full-length heterologous polypeptide as a percentage of total such heterologous polypeptide in prokaryotic host cells comprising:
- (a) culturing the host cells, which comprise nucleic acid encoding GreA or GreB protein, nucleic acid encoding the heterologous polypeptide, and one or more promoters for the nucleic acids;
- and
- (b) recovering the heterologous polypeptide from the cells or from cell culture medium, whereby the amount of full-length heterologous polypeptide produced by the process is increased as a percentage of total said heterologous polypeptide produced.
Dependencies are inferred from the claim text.