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Use of degenerate primers for polymerase chain reaction cloning and sequencing of the Lactococcus lactis subsp. lactis recA gene

1992; American Society for Microbiology; Volume: 58; Issue: 8 Linguagem: Inglês

10.1128/aem.58.8.2674-2678.1992

1098-5336

Patrick Duwat, S. Dusko Ehrlich, Alexandra Gruss,

Bacterial Genetics and Biotechnology

Two particularly well-conserved stretches in the RecA protein sequences were chosen as templates to synthesize degenerate oligonucleotides, which were used in polymerase chain reaction to amplify an internal recA DNA fragment of Lactococcus lactis subsp. lactis ML3. Using this fragment, we recovered and sequenced the entire lactococcal recA gene. The end of an open reading frame present upstream of the recA gene shows strong homology with formamidopyrimidine-DNA-glycosylase, a protein involved in DNA repair.