Isolation and expression in Escherichia coli of a Xanthomonas oryzae recA-like gene.
The recA gene from the bacterium Xanthomonas oryzae pv. oryzae (Xoo), a rice pathogen, was cloned based on its ability to complement DNA repair defects of Escherichia coli recA- mutants. The Xoo recA was localized to a 1.3-kb Sau3AI-XhoI fragment and, when cloned into pBR322, specifies increased methylmethanesulfonate and mitomycin C resistance to E. coli recA mutants and allows lambda red- gam- to plaque on an E. coli recA- host. An E. coli recA- strain harboring a plasmid containing the Xoo recA-like gene was shown to produce a 40-kDa protein which cross-reacted with an anti-E. coli RecA antibody. A similar molecular mass protein to RecA has been detected in several Xanthomonas pathovars using an anti-E. coli RecA antibody. Furthermore, the cloned Xoo recA was shown to hybridize to genomic DNA from various Xanthomonas pathovars, but not to genomic DNA from other bacteria species under high-stringency hybridization conditions. These results indicate the isolation of the Xoo recA gene.
Rabibhadana
S
S
Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok, Thailand.
Chamnongpol
S
S
Trempy
J E
JE
Ambulos
N P
NP
Mongkolsuk
S
S
D010582:Bacteriophage lambda / Q000502:physiology; D015139:Blotting, Southern; D015153:Blotting, Western; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000187:drug effects; D005816:Genetic Complementation Test; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D016685:Mitomycin / Q000494:pharmacology; D011938:Rec A Recombinases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D013045:Species Specificity; D010948:Viral Plaque Assay; D014974:Xanthomonas / Q000235:genetics*
D016685:Mitomycin; D008741:Methyl Methanesulfonate; D011938:Rec A Recombinases