Isolation and expression in Escherichia coli of a Xanthomonas oryzae recA-like gene

Gene. 1993 Sep 30;132(1):113-8. doi: 10.1016/0378-1119(93)90522-5.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacteriophage lambda / physiology
  • Blotting, Southern
  • Blotting, Western
  • Cloning, Molecular
  • Escherichia coli / drug effects
  • Genetic Complementation Test
  • Methyl Methanesulfonate / pharmacology
  • Mitomycin / pharmacology
  • Rec A Recombinases / genetics*
  • Rec A Recombinases / isolation & purification
  • Rec A Recombinases / metabolism
  • Species Specificity
  • Viral Plaque Assay
  • Xanthomonas / genetics*

Substances

  • Mitomycin
  • Methyl Methanesulfonate
  • Rec A Recombinases