Geisler U, Schumann W
Lehrstuhl für Genetik, Universität Bayreuth, FRG.
FEMS Microbiol Lett. 1993 Apr 15;108(3):251-4. doi: 10.1111/j.1574-6968.1993.tb06110.x.
A novel genetic procedure is described to identify stress genes in Bacillus subtilis by insertion mutagenesis. In addition, this method allows the rapid mapping of the mutation and the establishment of the DNA sequence of the gene impaired by the mutation. Small restriction fragments of chromosomal DNA of B. subtilis are inserted into a pBR322-based vector, the recombinant plasmids are transformed into B. subtilis, and integrants are selected which arise by recombination between the insert and its homologous region within the bacterial chromosome. About two dozen heat-, cold- and salt-sensitive mutants were isolated. Four mutations were mapped using PBS1 transduction, and the physiology of one salt-sensitive mutant was analysed.
描述了一种通过插入诱变鉴定枯草芽孢杆菌中应激基因的新遗传方法。此外，该方法能够快速定位突变并确定因突变而受损的基因的DNA序列。将枯草芽孢杆菌染色体DNA的小限制性片段插入基于pBR322的载体中，将重组质粒转化到枯草芽孢杆菌中，并选择通过插入片段与其在细菌染色体内的同源区域之间的重组而产生的整合体。分离出了大约二十多个对热、冷和盐敏感的突变体。使用PBS1转导对四个突变进行了定位，并分析了一个盐敏感突变体的生理学特性。
