Zagorec M, Steinmetz M
Laboratoire de Génétique des Microorganismes, Institut National Agronomique, Thiverval-Grignon, France.
J Gen Microbiol. 1991 Jan;137(1):107-12. doi: 10.1099/00221287-137-1-107.
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 Tn917PF1 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 beta-galactosidase.
转座子Tn917的工程变体已被广泛用于在枯草芽孢杆菌和其他革兰氏阳性细菌中获得插入突变和转录融合。我们开发了一种基于Tn917的新方法，可用于分离和鉴定因基因过表达产生的突变体。构建了一种Tn917变体，其一端附近含有一个强外向启动子，能够促进其插入靶点附近基因的转录。这个转座子命名为Tn917PF1，并在模型条件下进行了测试。分析了枯草芽孢杆菌的三个Tn917PF1突变体，其表型推测是由基因过表达导致的。结果表明它们的表型是由于转座子启动子的转录所致。在一个突变体中，启动子激活了一个deg基因，可能是degQ。另外两个突变体含有不同的插入片段，这些插入片段解密了一个编码β-半乳糖苷酶的枯草芽孢杆菌基因。
