Distinguishing between the extracellular DNases of Vibrio cholerae and development of a transformation system.
Vibrio cholerae is known to secrete DNase(s) into the extracellular environment. These proteins have been thought to be responsible for the difficulties in transforming this organism. In this work we demonstrate that the dns and xds genes differ and that their products are solely responsible for the extracellular DNase activity. By site-directed mutagenesis, strains have been constructed which are mutant in one or both genes. These strains have been assessed for their ability to be transformed with plasmid DNA and for their virulence in the infant mouse cholera model. DNase-deficient mutants can be readily transformed and the product of dns appears to be the more significant barrier. No effect on virulence was observed with the mutants.
Focareta
T
T
Department of Microbiology and Immunology, University of Adelaide, South Australia.
Manning
P A
PA
D000818:Animals; D015139:Blotting, Southern; D002771:Cholera / Q000382:microbiology; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D003851:Deoxyribonucleases / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D051379:Mice; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D015183:Restriction Mapping; D014169:Transformation, Bacterial; D014734:Vibrio cholerae / Q000201:enzymology / Q000235:genetics* / Q000472:pathogenicity
D004269:DNA, Bacterial; D003851:Deoxyribonucleases