Sammons R L, Slynn G M, Smith D A
Department of Genetics, University of Birmingham, UK.
J Gen Microbiol. 1987 Dec;133(12):3299-312. doi: 10.1099/00221287-133-12-3299.
A transposon Tn917 insertion between gerE and ilvB has identified a new developmental locus, gerM, in Bacillus subtilis. gerM96::Tn917 affects both sporulation and germination. DNA on either side of the transposon has been cloned and includes the previously cloned sdhC and gerE loci. gerE terminates 2.1 kb from the end of the transposon. The gerM96::Tn917 mutant is oligosporogenous, yielding approximately 1% of the number of wild-type heat resistant spores in liquid medium and 10% on solid medium. Six hours after the onset of sporulation alkaline phosphatase and glucose dehydrogenase levels were 90% and 7%, respectively, of those of the wild-type. At this time 50% of the mutant cells were still dividing. The occurrence of multiple polar septa and 'pygmy' cells suggested a block at stage II of sporulation. Following addition of germinants, mutant spores prepared on nutrient agar lost heat resistance normally but released slightly less dipicolinic acid than wild-type spores. They also showed only partial loss of optical density, associated with a phase-grey appearance and striations in the cortex suggesting partial degradation. Expression of the gerM gene was monitored by production of beta-galactosidase encoded by a promotorless lacZ gene fused to the gerM96::Tn917 insertion. It occurred 1.5-4 h after commencement of sporulation. Transcription was directed from a promoter on the gerE side of gerM and was unaffected by a mutation in the gerE gene.
转座子Tn917插入枯草芽孢杆菌中gerE和ilvB之间,已鉴定出一个新的发育位点gerM。gerM96::Tn917影响芽孢形成和萌发。转座子两侧的DNA已被克隆,包括先前克隆的sdhC和gerE位点。gerE在距转座子末端2.1 kb处终止。gerM96::Tn917突变体是少孢子形成的,在液体培养基中产生的野生型耐热孢子数量约为1%,在固体培养基中为10%。芽孢形成开始6小时后,碱性磷酸酶和葡萄糖脱氢酶水平分别为野生型的90%和7%。此时,50%的突变细胞仍在分裂。多个极向隔膜和“侏儒”细胞的出现表明在芽孢形成的II期受阻。添加萌发剂后,在营养琼脂上制备的突变体孢子正常丧失耐热性,但释放的吡啶二羧酸比野生型孢子略少。它们还仅部分丧失光密度,伴有相灰色外观和皮层条纹,表明部分降解。通过由与gerM96::Tn917插入融合的无启动子lacZ基因编码的β-半乳糖苷酶的产生来监测gerM基因的表达。它在芽孢形成开始后1.5 - 4小时出现。转录由gerM的gerE侧的启动子指导,并且不受gerE基因突变的影响。