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Growth, osmotic downshock resistance and differentiation of Bacillus subtilis strains lacking mechanosensitive channels

机译:缺乏机械敏感通道的枯草芽孢杆菌菌株的生长,抗渗透冲击和分化

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Previous work has shown that the mechanosensitive (MS) channel of large conductance (MscL) is essential for preventing lysis of Bacillus subtilis log phase cells upon a rapid, severe osmotic downshock. Growing cells of B. subtilis strains lacking MscL and one or more putative MS channel proteins of small conductance (YhdY, YkuT and YfkC) showed even higher sensitivity to an osmotic downshock. The effect was greatest for a strain lacking MscL and YkuT, and a strain lacking all four MS channel proteins had a similar phenotype. These defects were complemented by expression of either MscL or YkuT in trans. All MS channel mutant strains ultimately became resistant to osmotic downshock in stationary phase but at varying times, with mscL ykuT strains taking the longest time to become resistant. Expression of β-galactosidase from gene fusions to lacZ showed modest expression of ykuT and lower levels of expression of yhdY and yfkC when strains were grown in medium containing high salt. Sporulation of all MS channel mutant strains was normal, and the mutant spores germinated normally with l-alanine or dodecylamine.
机译:先前的研究表明,大电导(MscL)的机械敏感(MS)通道对于防止枯草芽孢杆菌对数期细胞在快速,严重的渗透下降冲击下溶解至关重要。缺乏MscL的枯草芽孢杆菌菌株的生长细胞以及一种或多种电导率较小的推定MS通道蛋白(YhdY,YkuT和YfkC)显示出对渗透压下降的更高敏感性。对于缺乏MscL和YkuT的菌株,效果最大,而缺乏所有四个MS通道蛋白的菌株具有相似的表型。这些缺陷可以通过反式表达MscL或YkuT来弥补。所有的MS通道突变菌株最终都在固定相中对渗透压下降产生了抗性,但是在不同的时间,mscL ykuT菌株花费了最长的时间才具有抗性。当菌株在高盐培养基中生长时,基因融合到lacZ的β-半乳糖苷酶表达显示ykuT适度表达,yhdY和yfkC表达较低。所有MS通道突变株的孢子形成均正常,并且突变孢子通常用1-丙氨酸或十二烷基胺萌发。

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