A genetic and molecular characterization of the recA gene from Staphylococcus aureus

Kenneth W Bayles, E. W. Brunskill, J. J. landolo, L. L. Hruska, S. Huang, P. A. Pattee, B. K. Smiley, R. E. Yasbin

Research output: Contribution to journalArticle

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Abstract

Previous studies have identified mutant strains of Staphylococcus aureus that have deficiencies in genetic recombination and DNA repair. Although these phenotypes were tentatively attributed to mutations within the S. aureus recA gene, experimental evidence to confirm this has never been reported. To characterize recA from S. aureus, we first isolated transposon insertion mutations that were in close proximity to the recA-like mutation (uus-568) in strain 112 UVS-1. This allowed for the mobilization of the uus-568 mutation into strain RN4220, the common laboratory strain of S. aureus. Next, using Bacillus subtilis recA as a probe, we cloned S. aureu recA and determined its nucleotide sequence. The deduced amino acid (aa) sequence of RecA contained 347 aa and was 74% identical to B. subtilis RecA. Using a cloned DNA fragment originating from within S. aureu recA, we then constructed a recA null mutant strain, designated KB 103, which exhibited the same phenotypic characteristics imposed by the uvs-56S mutation in the same background. Furthermore, genetic and physical mapping of S. aureu recA placed it in the same region as the uus-568 mutation. These data strongly suggest that these mutations represent different alleles of the same recA gene. RecA; recombination; DNA repair; Campbell integration; Cheo Box.

Original languageEnglish (US)
Pages (from-to)13-20
Number of pages8
JournalGene
Volume147
Issue number1
DOIs
StatePublished - Sep 15 1994

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Staphylococcus aureus
Molecular Biology
Mutation
Genes
Bacillus subtilis
DNA Repair
Genetic Recombination
Physical Chromosome Mapping
Recombinational DNA Repair
Insertional Mutagenesis
Amino Acid Sequence
Alleles
Phenotype
Amino Acids
DNA

ASJC Scopus subject areas

  • Genetics

Cite this

Bayles, K. W., Brunskill, E. W., landolo, J. J., Hruska, L. L., Huang, S., Pattee, P. A., ... Yasbin, R. E. (1994). A genetic and molecular characterization of the recA gene from Staphylococcus aureus. Gene, 147(1), 13-20. https://doi.org/10.1016/0378-1119(94)90033-7

A genetic and molecular characterization of the recA gene from Staphylococcus aureus. / Bayles, Kenneth W; Brunskill, E. W.; landolo, J. J.; Hruska, L. L.; Huang, S.; Pattee, P. A.; Smiley, B. K.; Yasbin, R. E.

In: Gene, Vol. 147, No. 1, 15.09.1994, p. 13-20.

Research output: Contribution to journalArticle

Bayles, KW, Brunskill, EW, landolo, JJ, Hruska, LL, Huang, S, Pattee, PA, Smiley, BK & Yasbin, RE 1994, 'A genetic and molecular characterization of the recA gene from Staphylococcus aureus', Gene, vol. 147, no. 1, pp. 13-20. https://doi.org/10.1016/0378-1119(94)90033-7
Bayles KW, Brunskill EW, landolo JJ, Hruska LL, Huang S, Pattee PA et al. A genetic and molecular characterization of the recA gene from Staphylococcus aureus. Gene. 1994 Sep 15;147(1):13-20. https://doi.org/10.1016/0378-1119(94)90033-7
Bayles, Kenneth W ; Brunskill, E. W. ; landolo, J. J. ; Hruska, L. L. ; Huang, S. ; Pattee, P. A. ; Smiley, B. K. ; Yasbin, R. E. / A genetic and molecular characterization of the recA gene from Staphylococcus aureus. In: Gene. 1994 ; Vol. 147, No. 1. pp. 13-20.
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abstract = "Previous studies have identified mutant strains of Staphylococcus aureus that have deficiencies in genetic recombination and DNA repair. Although these phenotypes were tentatively attributed to mutations within the S. aureus recA gene, experimental evidence to confirm this has never been reported. To characterize recA from S. aureus, we first isolated transposon insertion mutations that were in close proximity to the recA-like mutation (uus-568) in strain 112 UVS-1. This allowed for the mobilization of the uus-568 mutation into strain RN4220, the common laboratory strain of S. aureus. Next, using Bacillus subtilis recA as a probe, we cloned S. aureu recA and determined its nucleotide sequence. The deduced amino acid (aa) sequence of RecA contained 347 aa and was 74{\%} identical to B. subtilis RecA. Using a cloned DNA fragment originating from within S. aureu recA, we then constructed a recA null mutant strain, designated KB 103, which exhibited the same phenotypic characteristics imposed by the uvs-56S mutation in the same background. Furthermore, genetic and physical mapping of S. aureu recA placed it in the same region as the uus-568 mutation. These data strongly suggest that these mutations represent different alleles of the same recA gene. RecA; recombination; DNA repair; Campbell integration; Cheo Box.",
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