Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Enzymatic production of a protein or polypeptide
Patent
1983-07-25
1987-02-17
Wiseman, Thomas G.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Enzymatic production of a protein or polypeptide
4351723, 435317, 935 6, 935 29, 935 41, 935 48, 935 56, 935 72, C12P 2100, C12N 1500, C12N 100
Patent
active
046439692
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 (on an insertion site therefor) linked for transcriptional expression in reading phase with four functional fragments derived from the lipoprotein gene of E. coli. The plasmids further include a DNA sequence coding for a specific segment of the E. coli lac promoter-operator, which is positioned in the proper orientation for transcriptional expression of the desired polypeptide, as well as a separate functional E. coli lacI gene coding for the associated repressor molecule which can interact with the lac promoter-operator 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 promotor 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 promoter. The methods utilize such plasmids to introduce genetic capability into micro-organisms for the production of proteins, such as medically or commerically useful hormones, enzymes, immunogenic proteins, or intermediates therefor, but only in the presence of an appropriate inducer.
REFERENCES:
patent: 4237244 (1980-12-01), Cohen et al.
Masui et al, "Multipurpose Expression Cloning Vehicles in E. Coli", Date: 5/23/83.
Nakamura et al, The EMBO Journal, vol. 1, pp. 771-775, Jun. 1982.
Nakamura et al, Journal of Molecular and Applied Genetics, vol. 1, pp. 289-299, Jun. 1982.
Inouye Masayori
Masui Yoshihiro
Gresser J. M.
The Research Foundation of State University of New York
Wiseman Thomas G.
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