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Selective Inactivation of the Exonuclease Activity of Bacteriophage T7 DNA Polymerase by in Vitro Mutagenesis
Open accessAbstract
The 3' to 5' exonuclease activity of bacteriophage T7 DNA polymerase (gene 5 protein) can be inactivated selectively by reactive oxygen species. Differences in the enzymatic properties between the two forms are exploited to show by a chemical screen that modification of a histidine residue reduces selectively the exonuclease activity. In vitro mutagenesis of the histidine at residue 123, and of the neighboring residues, results in varying reduction of the exonuclease activity, including mutant enzymes that have no detectable exonuclease activity; as a consequence their polymerase activity is increased up to 9-fold. T7 phage containing the mutant genes have a greatly reduced burst size and demonstrate up to a 14-fold increase in the spontaneous mutation rate.
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Cited by
Initial sequencing and analysis of the human genome
International Human Genome Sequencing Consortium, Whitehead Institute for Biomedical Research, C. F. G. R., Lander, E. S., Linton, L. M., Birren, B., Nusbaum, C., Zody, M. C., Baldwin, J., Devon, K., Dewar, K., et al. (2001) · Nature · doi:10.1038/35057062 · Cited by 24,216
8-Hydroxyguanine, an abundant form of oxidative DNA damage, causes G-T and A-C substitutions.
Cheng, K. C., Cahill, D. S., Kasai, H., Nishimura, S., & Loeb, L. A. (1992) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)48474-8 · Cited by 1,844
DNA polymerase fidelity and the polymerase chain reaction.
Eckert, K. A., & Kunkel, T. A. (1991) · Genome Research · doi:10.1101/gr.1.1.17 · Cited by 448
Tabor, S., & Richardson, C. C. (1989) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.86.11.4076 · Cited by 288
Self-citationTsuchihashi, Z., & Brown, P. O. (1994) · Journal of Virology · doi:10.1128/jvi.68.9.5863-5870.1994 · Cited by 253
Metadata
- Authors
- Tabor, S., & Richardson, C. C.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 1989-04
- Volume
- 264
- Issue
- 11
- Pages
- 6447-6458
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W1551501575
- Cited by
- 255
- References
- 0
- FWCI
- 15.71
- Type
- journal article
- Language
- en
- Open access
- hybrid
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- Tabor, S., & Richardson, C. C. (1989). Selective Inactivation of the Exonuclease Activity of Bacteriophage T7 DNA Polymerase by in Vitro Mutagenesis. Journal of Biological Chemistry. https://doi.org/10.1016/s0021-9258(18)83369-5
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