Open AccessConstruction of a derivative of Tn917 containing an outward-directed promoter and its use in Bacillus subtilis
Author(s) -
Monique Zagorec,
Michel O. Steinmetz
Publication year - 1991
Publication title -
microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.019
H-Index - 179
eISSN - 1465-2080
pISSN - 1350-0872
DOI - 10.1099/00221287-137-1-107
Subject(s) - bacillus subtilis , transposable element , mutant , gene , biology , transcription (linguistics) , promoter , genetics , insertion , phenotype , gene expression , microbiology and biotechnology , bacteria , linguistics , philosophy
Summary: Engineered variants of the transposon Tn 917 have been widely used to obtain insertion mutations and transcriptional fusions in Bacillus subtilis and other Gram-positive bacteria. We have developed a novel Tn 917 -based methodology useful for isolation and characterization of mutants resulting from gene over-expression. A Tn 917 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 Tn 917PF1 , was tested in model conditions. Three Tn 917PF1 mutants of B. subtilis , with phenotypes presumed to result from gene over-expression, 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 β -galactosidase.