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枯草芽孢杆菌中一个多糖脱乙酰酶基因同源物(pdaB)对芽孢形成的特性研究

Characterization of a polysaccharide deacetylase gene homologue (pdaB) on sporulation of Bacillus subtilis.

作者信息

Fukushima Tatsuya, Tanabe Taichi, Yamamoto Hiroki, Hosoya Shigeo, Sato Tsutomu, Yoshikawa Hirofumi, Sekiguchi Junichi

机构信息

Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, Ueda, Nagano 386-8567, Japan.

出版信息

J Biochem. 2004 Sep;136(3):283-91. doi: 10.1093/jb/mvh151.

Abstract

The predicted amino acid sequence of Bacillus subtilis ybaN (renamed pdaB) exhibits high similarity to those of several polysaccharide deacetylases. Northern hybridization analysis with sporulation sigma mutants indicated that the pdaB gene is transcribed by EsigmaE RNA polymerase and negatively regulated by SpoIIID. The pdaB mutant was deficient in spore formation. Phase- and electron microscopic observation showed morphological changes of spores in late sporulation periods. The pdaB spores that had lost their viability were empty. Moreover, GFP driven by the promoter of the sspE gene was localized in the forespore compartment for the wild type, but was localized in both the mother cell and forespore compartments for phase-gray/dark forespores of the pdaB mutant. This indicates that GFP expressed in the forespores of the mutant leaks into the mother cells. Therefore, PdaB is necessary to maintain spores after the late stage of sporulation.

摘要

枯草芽孢杆菌ybaN(重命名为pdaB)的预测氨基酸序列与几种多糖脱乙酰酶的序列高度相似。用芽孢形成σ因子突变体进行的Northern杂交分析表明,pdaB基因由EσE RNA聚合酶转录,并受SpoIIID负调控。pdaB突变体在孢子形成方面存在缺陷。相差显微镜和电子显微镜观察显示,在芽孢形成后期孢子的形态发生了变化。失去活力的pdaB孢子是空的。此外,由sspE基因启动子驱动的绿色荧光蛋白(GFP)在野生型中定位于前芽孢区室,但在pdaB突变体的灰色/深色前芽孢的母细胞和前芽孢区室中均有定位。这表明在突变体前芽孢中表达的GFP会泄漏到母细胞中。因此,PdaB对于芽孢形成后期维持孢子是必需的。

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