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Replacement and amplification of bacterial genes with sequences altered in vitro.
Open accessAbstract
An efficient method for the replacement of chromosomal DNA by segments altered in vitro has been developed for bacteria. The method requires (i) a recombinant plasmid with a ColE1-like replicon and (ii) a strain defective in DNA polymerase I (polA), which is unable to replicate the plasmid extrachromosomally. This method is of general use since there are a number of suitable vectors and polA strains are available in both Escherichia coli and Salmonella typhimurium, the two most widely studied bacterial species. Using the method, we have constructed two chromosomal deletions in the chemotaxis gene region of S. typhimurium. In addition, plasmid sequences integrated into the chromosome have been amplified up to 30-fold by varying the concentration of ampicillin or tetracycline in the growth medium.
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Citations
Cited by
Link, A. J., Phillips, D., & Church, G. M. (1997) · Journal of Bacteriology · doi:10.1128/jb.179.20.6228-6237.1997 · Cited by 874
Use of Bacteriophage λ Recombination Functions To Promote Gene Replacement in Escherichia coli
Murphy, K. C. (1998) · Journal of Bacteriology · doi:10.1128/jb.180.8.2063-2071.1998 · Cited by 573
Ried, J. L., & Collmer, A. (1987) · Gene · doi:10.1016/0378-1119(87)90127-2 · Cited by 351
Codon Pair Utilization Biases Influence Translational Elongation Step Times
Irwin, B., Heck, J. D., & Hatfield, G. W. (1995) · Journal of Biological Chemistry · doi:10.1074/jbc.270.39.22801 · Cited by 166
Construction and characterization of an Escherichia coli strain with a uncI mutation
Gay, N. J. (1984) · Journal of Bacteriology · doi:10.1128/jb.158.3.820-825.1984 · Cited by 89
Metadata
- Authors
- Gutterson, N. I., & Koshland, D. E.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 1983-08
- Volume
- 80
- Issue
- 16
- Pages
- 4894-4898
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W2003511537
- Cited by
- 180
- References
- 0
- FWCI
- 10.35
- Type
- journal article
- Language
- en
- Open access
- green
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- Gutterson, N. I., & Koshland, D. E. (1983). Replacement and amplification of bacterial genes with sequences altered in vitro. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.80.16.4894
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