Molecular cloning and characterization of the recA gene of Methylomonas clara and construction of recA deficient mutant.
The recA gene of the methylotrophic bacterium Methylomonas clara has been isolated from a genomic library by hybridization with the Escherichia coli recA gene. Its complete nucleotide sequence consists of 1029 bp encoding a polypeptide of 342 amino acids. Nucleotide sequence analysis of the M. clara recA gene revealed extensive homologies to recA genes from E. coli and Pseudomonas aeruginosa. Part of the physiological activity of the M. clara RecA protein has become evident in that E. coli recA mutant HB101 is complemented. The cloned recA gene has been modified in vitro by site-specific mutagenesis and by insertion of a kanamycin-resistance gene cassette into the recA coding sequence. M. clara recA mutants were obtained by replacement of the active recA gene by an in-vitro inactivated gene copy.
Lehrstuhl für Allgemeine Botanik, Ruhr-Universität Bochum, Federal Republic of Germany.
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Ridder
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Marquardt
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Esser
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D019605:Euryarchaeota / Q000235:genetics* / Q000528:radiation effects; D015870:Gene Expression; D005798:Genes, Bacterial; D007613:Kanamycin Resistance; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011938:Rec A Recombinases / Q000235:genetics*; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014466:Ultraviolet Rays
D004269:DNA, Bacterial; D011938:Rec A Recombinases