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. 1989 May;171(5):2614–2618. doi: 10.1128/jb.171.5.2614-2618.1989

Exchange of chromosomal and plasmid alleles in Escherichia coli by selection for loss of a dominant antibiotic sensitivity marker.

C B Russell 1, F W Dahlquist 1
PMCID: PMC209942  PMID: 2651409

Abstract

Transfer of an allele from a donor DNA to a recipient DNA molecule was selected by the loss of a dominant conditional lethal mutation previously incorporated ito the gene of interest in the recipient DNA. Both the Escherichia coli chromosome and plasmids carrying E. coli genes were used successfully as donor molecules. Recipient molecules for these exchanges were constructed in vitro by using the rpsL gene, which confers sensitivity to streptomycin, to replace segments of specific E. coli genes located either on multicopy plasmids or in the E. coli chromosome. Plasmids carrying such replacements were capable of acquiring chromosomal alleles of the gene(s) of interest, and strains carrying rpsL replacements in the chromosome were capable of acquiring plasmid-encoded alleles at the sight of the rpsL replacement. In both situations, these allele transfers resulted in loss of the rpsL gene from the recipient DNA molecule. The desired transfer events constituted a large percentage of these events, which gave rise to viable colonies when appropriate donor-recipient pairs were subjected to streptomycin selection. Thus, this is a useful approach for transferring alleles of interest from plasmids to the E. coli chromosome and vice versa.

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Selected References

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

  1. Biek D. P., Cohen S. N. Identification and characterization of recD, a gene affecting plasmid maintenance and recombination in Escherichia coli. J Bacteriol. 1986 Aug;167(2):594–603. doi: 10.1128/jb.167.2.594-603.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Davies J., Nomura M. The genetics of bacterial ribosomes. Annu Rev Genet. 1972;6:203–234. doi: 10.1146/annurev.ge.06.120172.001223. [DOI] [PubMed] [Google Scholar]
  3. Dean D. A plasmid cloning vector for the direct selection of strains carrying recombinant plasmids. Gene. 1981 Oct;15(1):99–102. doi: 10.1016/0378-1119(81)90108-6. [DOI] [PubMed] [Google Scholar]
  4. Gutterson N. I., Koshland D. E., Jr Replacement and amplification of bacterial genes with sequences altered in vitro. Proc Natl Acad Sci U S A. 1983 Aug;80(16):4894–4898. doi: 10.1073/pnas.80.16.4894. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  6. LEDERBERG J. Streptomycin resistance; a genetically recessive mutation. J Bacteriol. 1951 May;61(5):549–550. doi: 10.1128/jb.61.5.549-550.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Liljeström P., Pirhonen M., Palva E. T. In vivo transfer of chromosomal mutations onto multicopy plasmids by transduction with bacteriophage P1. Gene. 1985;40(2-3):241–246. doi: 10.1016/0378-1119(85)90046-0. [DOI] [PubMed] [Google Scholar]
  8. Mutoh N., Simon M. I. Nucleotide sequence corresponding to five chemotaxis genes in Escherichia coli. J Bacteriol. 1986 Jan;165(1):161–166. doi: 10.1128/jb.165.1.161-166.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Oka A., Sugisaki H., Takanami M. Nucleotide sequence of the kanamycin resistance transposon Tn903. J Mol Biol. 1981 Apr 5;147(2):217–226. doi: 10.1016/0022-2836(81)90438-1. [DOI] [PubMed] [Google Scholar]
  10. Park C., Hazelbauer G. L. Transfer of chromosomal mutations to plasmids via Hfr-mediated conduction. J Bacteriol. 1986 Jan;165(1):312–314. doi: 10.1128/jb.165.1.312-314.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Parkinson J. S. Complementation analysis and deletion mapping of Escherichia coli mutants defective in chemotaxis. J Bacteriol. 1978 Jul;135(1):45–53. doi: 10.1128/jb.135.1.45-53.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Russell C. B., Thaler D. S., Dahlquist F. W. Chromosomal transformation of Escherichia coli recD strains with linearized plasmids. J Bacteriol. 1989 May;171(5):2609–2613. doi: 10.1128/jb.171.5.2609-2613.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Silverman M., Simon M. Identification of polypeptides necessary for chemotaxis in Escherichia coli. J Bacteriol. 1977 Jun;130(3):1317–1325. doi: 10.1128/jb.130.3.1317-1325.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]

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