Construction of an Azospirillum brasilense Sp7 recA mutant |
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Authors: | Chris Croes Jos Vanderleyden Kris Michiels |
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Affiliation: | (1) F.A. Janssens Memorial Laboratory for Genetics, Catholic University of Leuven, B-3030 Heverlee, Belgium |
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Abstract: | Summary Cosmid clones encoding the recA gene of Azospirillum brasilense were isolated by intergeneric complementation of an Escherichia coli recA mutant. Site-directed Tn5 mutagenesis and subcloning of one complementing cosmid clone allowed us to localize the A. brasilense recA gene on a 1.2 kb DNA fragment. One Tn5 insertion that inactivates the cloned recA gene was crossed into the chromosome of A. brasilense by marker exchange. The resulting A. brasilense recA mutant showed increased sensitivity to the DNA methylating agent methyl methanesulfonate and to ultraviolet light and had at least one hundredfold reduced recombinational activity compared to the parent strain. |
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Keywords: | Azospirillum recA Homologous recombination Transposon Tn5 mutagenesis |
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