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PMID: 8294027 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Overproduction of the toxic protein, bovine pancreatic DNaseI, in Escherichia coli using a tightly controlled T7-promoter-based vector.

Gene ·Vol. 136 ·No. 1-2 ·1993-12-22 ·Pages 337-40

Doherty AJ, Connolly BA, Worrall AF

Abstract

A synthetic gene coding for bovine pancreatic DNaseI has been cloned under the control of a T7 promoter present on the plasmid pET11. This construct yields a stable Escherichia coli transformant only when transcription from this promoter is tightly controlled. Production of recombinant DNaseI (reDNaseI) is achieved by infection of the cells with a mutant lambda phage, CE6, which carries the gene encoding T7 RNA polymerase. Induced bacterial cultures yield in excess of 2 mg per litre of reDNaseI after purification.

MeSH Terms
Animals Bacteriophage T7/genetics Cattle Cloning, Molecular DNA-Directed RNA Polymerases/genetics Deoxyribonuclease I/genetics Escherichia coli/genetics Genes, Synthetic Pancreas/enzymology Plasmids Promoter Regions, Genetic Viral Proteins
Chemicals
Viral Proteins bacteriophage T7 RNA polymerase DNA-Directed RNA Polymerases Deoxyribonuclease I
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Doherty A J
Department of Biochemistry, University of Southampton, UK.
Connolly B A
Worrall A F
Article Info
Journal
Gene
Abbr.
Gene
ISSN
0378-1119
Published
1993-12-22
Pages
337-40
Language
English
Region
Netherlands
NLM ID
7706761
Subset
IM
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