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A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes.
Open accessAbstract
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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Cited by
A General Method for Site-specific Incorporation of Unnatural Amino Acids into Proteins
Noren, C. J., Anthony-Cahill, S. J., Griffith, M. C., & Schultz, P. G. (1989) · Science · doi:10.1126/science.2649980 · Cited by 1,019
The ZnuABC high‐affinity zinc uptake system and its regulator Zur in Escherichia coli
Patzer, S. I., & Hantke, K. (1998) · Molecular Microbiology · doi:10.1046/j.1365-2958.1998.00883.x · Cited by 490
Characterization of the ferrous iron uptake system of Escherichia coli
Kammler, M., Schön, C., & Hantke, K. (1993) · Journal of Bacteriology · doi:10.1128/jb.175.19.6212-6219.1993 · Cited by 460
Ratliff, M., Zhu, W., Deshmukh, R., Wilks, A., & Stojiljkovic, I. (2001) · Journal of Bacteriology · doi:10.1128/jb.183.21.6394-6403.2001 · Cited by 221
Stojiljkovic, I., & Hantke, K. (1994) · Molecular Microbiology · doi:10.1111/j.1365-2958.1994.tb00465.x · Cited by 205
Metadata
- Authors
- Tabor, S., & Richardson, C. C.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 1985-02
- Volume
- 82
- Issue
- 4
- Pages
- 1074-1078
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W2068518785
- Cited by
- 3,148
- References
- 0
- FWCI
- 33.24
- Type
- journal article
- Language
- en
- Open access
- green
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- Tabor, S., & Richardson, C. C. (1985). A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.82.4.1074
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