Exogenous phage recombinase-independent inactivation of chromosomal genes in Yersinia enterocolitica.
Characterization of newly identified genes is necessary to understand their functions. Phenotypic characterization of isogenic mutants provides good understanding of the functions of the genes in wild type strains. In the present study, we report the use of linear dsDNA as a substrate for homologous recombination in Yersinia enterocolitica. A double-stranded linear recombinant DNA (LRD) containing an antibiotic resistance gene flanked by homologous regions to the target gene was created. Transformation of this LRD into Y. enterocolitica led to the replacement of targeted loci with antibiotic resistance gene. Using this strategy, two chromosomal genes namely urease C (ureC) and hemophore A (hasA) were disrupted in three strains of Y. enterocolitica. These recombinations were independent of the EPR functions. This is the first report of EPR-independent inactivation of chromosomal genes in Y. enterocolitica strains.
Microbial Pathogenicity Laboratory, University of Delhi South Campus, Benito Juarez Road, New Delhi 110 021, India. Electronic address: maheshsdhar@gmail.com.
Mahesh S
MS
Dhar
Pradeep
P
Kumar
Jugsharan S
JS
Virdi
D001426:Bacterial Proteins / Q000235:genetics*; D001435:Bacteriophages / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics; D020868:Gene Silencing; D018390:Gene Targeting / Q000379:methods*; D005798:Genes, Bacterial; D009154:Mutation; D045522:Recombinases / Q000235:genetics; D011995:Recombination, Genetic; D015008:Yersinia enterocolitica / Q000235:genetics*
D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D004274:DNA, Recombinant; D045522:Recombinases; C474041:ureC protein, Yersinia enterocolitica
Homologous recombination; Linear recombinant DNA; Yersinia enterocolitica