Construction and properties of Tn917-lac, a transposon derivative that mediates transcriptional gene fusions in Bacillus subtilis.
Construction and properties of Tn917-lac, a transposon derivative that mediates transcriptional gene fusions in Bacillus subtilis.
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Tn917-lac（一种介导枯草芽孢杆菌转录基因融合的转座子衍生物）的构建和特性。
DOI：
10.1073/pnas.83.1.140
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发表时间：
1986
影响因子：
11.1
通讯作者：
Youngman,PJ
中科院分区：
文献类型：
--
作者：
Perkins,JB;Youngman,PJ
构建了Tn 917的衍生物，称为Tn 917-lac，其能够产生将枯草芽孢杆菌染色体基因的转录物连接到大肠杆菌lacZ基因的编码序列的融合体。Tn 917-lac的两个独立插入到B的gltA基因中，一个插入到trpE基因中（在trpEDCFBA操纵子中）。详细研究了Tn 917-lac-mediated的转录融合，结果证实Tn 917-lac-mediated的转录融合产生的β-半乳糖苷酶水平准确地反映了中断基因的调节表达。为了促进这些研究，开发了一种程序，允许分析Tn 917-lac-mediated融合在部分二倍体中，其中插入突变由中断基因的完整拷贝补充。已知Tn 917在代表三种完全不同的革兰氏阳性属（链球菌属、芽孢杆菌属和葡萄球菌属）的细菌中有效地起作用，并且已知Tn 917在其插入细菌基因组中时显示出相对高度的随机性，使得Tn 917-lac可能用于鉴定和研究广泛范围的革兰氏阳性物种中的多种调节基因。
A derivative of Tn917 was constructed, referred to as Tn917-lac, which is capable of generating fusions that connect the transcripts of Bacillus subtilis chromosomal genes to the coding sequence of the lacZ gene of Escherichia coli. Two independent insertions of Tn917-lac into the gltA gene and one insertion into the trpE gene (in the trpEDCFBA operon) of B. subtilis were studied in detail, and the results confirmed that Tn917-lac-mediated transcriptional fusions produce levels of beta-galactosidase that reflect accurately the regulated expression of interrupted genes. To facilitate these studies, a procedure was developed that permits the analysis of Tn917-lac-mediated fusions in partial diploids where insertional mutations are complemented by an intact copy of the interrupted genes. Tn917 is known to function efficiently in bacteria representing three quite different Gram-positive genera (Streptococcus, Bacillus, and Staphylococcus) and is known to display a relatively high degree of randomness in its insertions into bacterial genomes, making it likely that Tn917-lac will be useful for the identification and study of many kinds of regulated genes in a wide range of Gram-positive species.