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. 1985 Feb;82(4):1074–1078. doi: 10.1073/pnas.82.4.1074

A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes.

S Tabor, C C Richardson
PMCID: PMC397196  PMID: 3156376

Abstract

The RNA polymerase gene of bacteriophage T7 has been cloned into the plasmid pBR322 under the inducible control of the lambda PL promoter. After induction, T7 RNA polymerase constitutes 20% of the soluble protein of Escherichia coli, a 200-fold increase over levels found in T7-infected cells. The overproduced enzyme has been purified to homogeneity. During extraction the enzyme is sensitive to a specific proteolysis, a reaction that can be prevented by a modification of lysis conditions. The specificity of T7 RNA polymerase for its own promoters, combined with the ability to inhibit selectively the host RNA polymerase with rifampicin, permits the exclusive expression of genes under the control of a T7 RNA polymerase promoter. We describe such a coupled system and its use to express high levels of phage T7 gene 5 protein, a subunit of T7 DNA polymerase.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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