THE ilvBC-leu OPERON OF BACILLUS SUBTILIS11Supported in part by grant GM33152 from the National Institute of General Medical Sciences.

~~~~~~~~~~~ ~~The most common effect of transposon insertion is the inactivation of genes. However, in some cases, transposons can activate in cis the expression of genes in the neighbourhood of their integration site. We previously described an insertion of the transposon Tn917 into the Bacillus subtilis sacXY locus. smX and sacY encode respectively a negative and a positive regulator involved in induction by sucrose of the exoenzyme levansucrase. Data in this paper show that the Tn917 insertion had two effects: it inactivated sacXand it increased the transcription of sacX The latter effect involved one or several elements internal to the transposon.

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Construction of a derivative of Tn917 containing an outward-directed promoter and its use in Bacillus subtilis

Zagorec¹,

Steinmetz²

1991

Microbiology

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Engineered variants of the transposon Tn917 have been widely used to obtain insertion mutations and transcriptional fusions in Bacillus subtilis and other Gram-positive bacteria. We have developed a novel Tn917-based methodology useful for isolation and characterization of mutants resulting from gene over-expression. A Tn917 variant was constructed which contains a strong out-facing promoter near one end, able to promote transcription of genes in the vicinity of its insertion target. This transposon, designated Tn917PF1, was tested in model conditions. Three Tn917PFI mutants of B. subtilis, with phenotypes presumed to result from gene overexpression, were analysed. Their phenotypes were shown to be due to transcription from the transposon promoter. In one mutant the promoter activated a deg gene, probably degQ. The other two contained different insertions decryptifying a B. subtilis gene encoding P-galactosidase.

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THE ilvBC-leu OPERON OF BACILLUS SUBTILIS11Supported in part by grant GM33152 from the National Institute of General Medical Sciences.

Cited by 6 publications

References 5 publications

Temperate Bacteriophages of Bacillus subtilis

Temperate Bacteriophages of Bacillus subtilis

Dual effect of a Tn917 insertion into the Bacillus subtilis sacX gene

Construction of a derivative of Tn917 containing an outward-directed promoter and its use in Bacillus subtilis

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