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Current Microbiology

, Volume 23, Issue 3, pp 123–129 | Cite as

Cloning and expression inEscherichia coli of arecA-like gene fromZymomonas mobilis

  • T. Karunakaran
  • P. Gunasekaran
Article

Abstract

Intergeneric complementation ofEscherichia coli recA mutants was used to identify recombinant plasmids, within a genomic library derived fromZymomonas mobilis, that carryZ. mobilis recA-like gene. Screening of 1100 individualE. coli strains revealed four clones expressing therecA+ character. On restriction analysis, all four recombinant plasmids were found to be related and to exhibit a common 6.7-kb fragment. Consequently, one of the four recombinant plasmids, pZR27, was selected for further characterization. When introduced intoE. coli recA mutants, pZR27 restored resistance to methyl methane sulfonate, mitomycin-C, and UV irradiation, as well as recombination proficiency when measured by standard Hfr-mediated conjugation. The clonedrecA-like gene also restored the spontaneous and mitomycin-C-induced phage production. The origin of the insert in pZR27 from the chromosome ofZ. mobilis was confirmed by Southern transfer and DNA hybridization. However, no homology was found between therecA ofE. coli andZ. mobilis chromosomal insert DNA. TheZ. mobilis recA-like gene also encoded a major polypeptide of 38-kDa on SDS-PAGE.

Keywords

Recombination Recombinant Plasmid Restriction Analysis Genomic Library Methane Sulfonate 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag New York Inc. 1991

Authors and Affiliations

  • T. Karunakaran
    • 1
  • P. Gunasekaran
    • 1
  1. 1.Department of Microbiology and Microbial Technology, School of Biological SciencesMadurai Kamaraj UniversityMaduraiIndia

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