Construction of a slime negative transposon mutant in Staphylococcus epidermidis using the Enterococcus faecalis transposon Tn917.
The slime-producing Staphylococcus epidermidis strain sensu strictu CNS23 was transformed by protoplast transformation with the plasmid pTV1 which carries transposon Tn 917. Using this transposon mutagenesis system we obtained the Tn917-inserted mutant CT512, which has lost the ability to produce slime. A single insertion of the transposon Tn917 into the chromosome of CT512 could be detected by Southern hybridization. This mutant showed a significantly higher stability concerning its slime-negative phenotype compared with spontaneous slime-negative mutants of S. epidermidis strain CNS23. In slime-ELISA no slime-associated antigen could be detected in extracts of the transposon mutant. Compared to slime-positive S. epidermidis strains, CT512 lacked in accumulative growth in microtiter tube test.
Institut für Medizinische Mikrobiologie und Immunologie, Universität Hamburg, Universitätsklinikum, F.R.G.
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D004251:DNA Transposable Elements; D013293:Enterococcus faecalis / Q000235:genetics*; D006801:Humans; D016296:Mutagenesis; D010957:Plasmids; D011135:Polysaccharides, Bacterial / Q000096:biosynthesis / Q000235:genetics*; D013212:Staphylococcus epidermidis / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity; D014170:Transformation, Genetic; D014774:Virulence / Q000235:genetics
D004251:DNA Transposable Elements; D011135:Polysaccharides, Bacterial