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The htpG gene of Bacillus subtilis belongs to class III heat shock genes and is under negative control.

机译:枯草芽孢杆菌的htpG基因属于III类热休克基因处于负调控下。

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摘要

We show that the htpG gene of Bacillus subtilis is induced by heat, as has been reported for the Escherichia coli homolog. Analysis of different mutants revealed that the htpG gene belongs to class III heat shock genes in B. subtilis. An about 10-fold induction after thermal upshock was found at the levels of both transcription and translation, and this induction resulted from enhanced synthesis of mRNA. By primer extension, we identified one potential transcription start site immediately downstream of a putative sigmaA-dependent promoter which became activated after thermal upshift. Northern blot analysis revealed that htpG is part of a monocistronic transcriptional unit. An operon fusion where the complete region between htpG and its upstream gene was fused to the bgaB reporter gene accurately reflected htpG expression. Analysis of this fusion revealed that, in contrast to other class III heat shock genes, htpG was not induced by osmotic upshock, by ethanol, or by oxygen limitation, suggesting that it belongs to a subgroup within class III. Deletion of the region upstream of the putative promoter resulted in an enhanced basal level of htpG expression, but the 10-fold induction was retained, suggesting that the upstream sequences are involved in the regulation of expression in the absence of heat shock.
机译:我们显示,枯草芽孢杆菌的htpG基因是由热诱导的,如已报道的大肠杆菌同源物。对不同突变体的分析表明,htpG基因属于枯草芽孢杆菌中的III类热激基因。在热冲击后,在转录和翻译的水平上均发现了约10倍的诱导,而这种诱导是由于mRNA合成增强所致。通过引物延伸,我们确定了一个潜在的转录起始位点,紧接一个假定的sigmaA依赖性启动子的下游,该启动子在热升档后被激活。 Northern印迹分析表明,htpG是单顺反子转录单位的一部分。操纵子融合体,其中htpG及其上游基因之间的完整区域与bgaB报告基因融合,可准确反映htpG的表达。对该融合蛋白的分析表明,与其他III类热休克基因相反,htpG不受渗透压,乙醇或氧气限制的诱导,表明它属于III类内的一个亚组。推定的启动子上游区域的删除导致htpG表达的基础水平提高,但保留了10倍的诱导,这表明在没有热激的情况下上游序列参与了表达的调节。

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