Varon M, Fuchs N, Monosov M, Tolchinsky S, Rosenberg E
Department of Molecular Microbiology and Biotechnology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Ramat Aviv, Israel.
Antimicrob Agents Chemother. 1992 Oct;36(10):2316-21. doi: 10.1128/AAC.36.10.2316.
Transposition of TnV and Tn5lac into Myxococcus xanthus yielded 8,381 kanamycin-resistant mutants that were tested for antibiotic TA production. Twenty-four of the mutants were nonproducers of TA (less than 0.4 ng/ml), and 3 produced a higher level (2.5 micrograms/ml) than the parent strain (1.5 micrograms/ml). For most of the strains, there was 100% cotransduction between kanamycin resistance and the altered TA phenotype. Southern blot analysis of restriction digests of the mutant DNA indicated that the transposons were inserted at different sites on the M. xanthus chromosome. The TA genes were mapped by cotransduction between pairs of mutants following replacement of the initial insert of one of the pair with the tetracycline resistance transposon Tn5-132. Nine of the 13 nonproducers tested were linked over a 36-kb stretch of the chromosome. There was no linkage between one of the overproducers and any of the nonproducers tested.
将TnV和Tn5lac转座到黄色粘球菌中产生了8381个对卡那霉素有抗性的突变体,对这些突变体进行了抗生素TA产生的检测。其中24个突变体不产生TA(低于0.4纳克/毫升),3个突变体产生的TA水平(2.5微克/毫升)高于亲本菌株(1.5微克/毫升)。对于大多数菌株,卡那霉素抗性与改变的TA表型之间存在100%的共转导。对突变体DNA的限制性消化产物进行Southern印迹分析表明,转座子插入到了黄色粘球菌染色体的不同位点。在用四环素抗性转座子Tn5 - 132替换其中一对突变体中的初始插入片段后,通过对成对突变体之间的共转导来绘制TA基因图谱。在检测的13个不产生TA的突变体中,有9个在染色体上36千碱基对的区域内连锁。其中一个过量产生TA的突变体与所检测的任何一个不产生TA的突变体之间没有连锁关系。