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US 6,284,488 B1Granted
Protein expression system
AssigneeCelltech Therapeutics Limited
InventorsAndrew Neil Charles Weir, Andrew Mountain
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Abstract
A method for expressing recombinant molecules in bacterial hosts in a defined medium in the absence of antibiotic selection is disclosed. The method uses an expression vector comprising a regulatable promoter by which the production of foreign proteins may be controlled during the growth phase of the culture, and origin of replication maintaining medium vector copy number and a transcriptional promoter.
Claims11
A method for producing a heterologous protein in a bacterial host cell, comprising:
- a) transforming a bacterial host cell with an expression vector comprising a single origin of replication, a regulatable promoter, a transcription unit encoding the heterologous polypeptide, and a transcriptional terminator;
- b) expressing the heterologous protein in the absence of an inducer so that the single origin of replication maintains a vector copy number between 6 and 50;
- and
- c) culturing the host cell in defined media in absence of antibiotic selection.
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