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A recombinase A-deficient strain of Actinobacillus actinomycetemcomitans constructed by insertional mutagenesis using
Keith P Mintz1, Catherine Brissette, Paula M Fives-Taylor
1Department of Microbiology and Molecular Genetics, The Markey Center for Molecular Genetics, College of Medicine, University of Vermont, Burlington 05405, USA.
Abstract:
The gene coding for recA in the oral pathogen Actinobacillus actinomycetemcomitans SUNY 465 was cloned and sequenced. The DNA sequence coded for a 352-amino acid protein that was homologous to RecA of a variety of bacterial species. A derivative of a non-replicating mobilizable plasmid was constructed for directed mutagenesis in A. actinomycetemcomitans. A recA-deficient strain of A. actinomycetemcomitans was developed by homologous recombination of an internal recA fragment contained on the mobilizable suicide vector. The recA mutant strain was more sensitive to UV radiation and showed a reduced recombinatorial proficiency than the isogenic parent strain. These data suggest that recA of A. actinomycetemcomitans SUNY 465 is involved in the repair of DNA damage caused by UV irradiation and homologous recombination as determined for other bacteria.