Novel cloning vehicles for polypeptide expression in microbial h

Details Novel cloning vehicles for polypeptide expression in microbial h Novel cloning vehicles for polypeptide expression in microbial h

1982-05-14

1989-09-05

Martinell, James

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Enzymatic production of a protein or polypeptide

4352523, 43525233, 435320, 4351723, 935 38, 935 39, 935 43, 935 44, 935 66, 935 72, 935 73, C12P 2100, C12N 120, C12N 1500

Patent

active

048638552

ABSTRACT:
Methods and compositions are provided for regulated expression of polypeptides in transformed bacterial hosts. A novel class of plasmid cloning vehicles includes a DNA sequence coding for the desired polypeptide (or an insertion site therefor) linked for transcriptional expression in reading phase with one or more functional fragments derived from an outer membrane protein gene of a Gram-negative bacterium. The plasmids also include an inducible promoter sequence positioned in the proper orientation for transcriptional expression of the desired polypeptide, as well as a separate DNA sequence coding for a repressor molecule which can interact with the inducible promoter to prevent transcription therefrom. Expression of the desired polypeptide is under the control of both the constitutive promoter and the inducible promoter, although transcription from either promoter is normally blocked by the repressor molecule. However, the repressor can be selectively inactivated by means of an inducer molecule to permit transcriptional expression of the desired polypeptide from both promoters. The methods utilize such plasmids to introduce genetic capability into micro-organisms for the production of proteins, such as medically or commercially useful hormones, enzymes, immunogenic proteins, or intermediates therefor, but only in the presence of an appropriate inducer.

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