Introduction and expression of the Streptococcus faecalis transposon Tn916 into Bacillus thuringiensis subsp. israelensis and its use in comobilizing the Staphylococcus aureus resistance plasmid pC194

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1987
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Naglich, Joseph
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Robert E. Andrews
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Microbiology
Abstract

The Staphylococcus aureus plasmid, pC194, was introduced into Bacillus thuringiensis subsp. israelensis using the Streptococcus faecalis transposon Tn916 as a mobilizing agent. The conjugative S. faecalis transposon Tn916 was introduced into B. thuringiensis subsp. israelensis by filter matings with S. faecalis. B. thuringiensis transconjugants resistant to tetracycline were detected at a frequency of approximately 7.0 x10[superscript] -7 per recipient cell when filter matings were performed, but tetracycline-resistant transconjugants were not observed when a broth-mating protocol was used. The tetracycline resistance phenotype was not lost during serial passage of subsp. israelensis without antibiotic selection. Southern hybridization analysis revealed that Tn916 had inserted into several different sites on the B. thuringiensis subsp. israelensis chromosome. Conjugation-like movement of Tn916 was demonstrated when tetracycline-resistant B. thuringiensis transconjugants were mated with isogenic recipients. Transfer of pC194 only occurred when B. thuringiensis subsp. israelensis was mated with a B. subtilis donor containing pC194 and Tn916. B. thuringiensis transconjugants resistant to chloramphenicol and tetracycline were detected at a frequency of 1.96 x 10[superscript] -6 per recipient cell when filter matings were performed. Transconjugants resistant to chloramphenicol were not observed when a broth mating protocol was used. The tetracycline resistance phenotype was maintained during serial passage of B. thuringiensis without antibiotic selection, whereas the chloramphenicol resistance phenotype was not. Southern hybridization analysis revealed that Tn916 had inserted into several different sites on the B. thuringiensis chromosome, and pC194 was introduced as an autonomous plasmid into the B. thuringiensis genome.

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Thu Jan 01 00:00:00 UTC 1987