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首页> 外文期刊>Journal of bacteriology >Effect of a 20-kilodalton protein from Bacillus thuringiensis subsp. israelensis on production of the CytA protein by Escherichia coli.
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Effect of a 20-kilodalton protein from Bacillus thuringiensis subsp. israelensis on production of the CytA protein by Escherichia coli.

机译：苏云金芽孢杆菌亚种中20公斤蛋白质的作用以色列通过大肠杆菌生产CytA蛋白。

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CytA, a 27-kDa cytolytic crystal protein of Bacillus thuringiensis subsp. israelensis, is produced only at very low levels by recombinant Escherichia coli cells unless a 20-kDa B. thuringiensis subsp. israelensis protein is also present (K. M. McLean and H. R. Whiteley, J. Bacteriol. 169:1017-1023, 1987; L. F. Adams, J. E. Visick, and H. R. Whiteley, J. Bacteriol. 171:521-530, 1989). However, the data reported here demonstrate that the 20-kDa protein is not required for high-level CytA production in E. coli strains carrying mutations in rpoH, groEL, or dnaK, all of which affect the proteolytic ability of the cells. The 20-kDa protein also increases the amount of CryIVD (another B. thuringiensis subsp. israelensis crystal protein) and LacZX90 (a mutant of beta-galactosidase) made by E. coli. The latter phenomenon is attributable to an increase in the half-life of LacZX90, suggesting that the 20-kDa protein may stabilize this protein. The effect of the 20-kDa protein was also examined in vitro and in a T7 RNA polymerase expression system, and the possible significance of these results for the timing of proteolysis and of 20-kDa protein activity is discussed. Finally, the ability of a single antibody to coimmunoprecipitate CytA and the 20-kDa protein from E. coli extracts provides evidence for a protein-protein interaction that may be related to the mechanism of action of the 20-kDa protein.
机译：CytA，苏云金芽孢杆菌亚种的27 kDa细胞溶解晶体蛋白。除非有20 kDa的苏云金芽孢杆菌亚种，否则重组大肠杆菌细胞只能产生极少量的以色列。以色列蛋白质也存在（K.M.McLean和H.R.Whiteley，J.Bacteriol.169：1017-1023，1987; L.F.Adams，J.E.Visick，和H.R.Whiteley，J.Bacteriol.171：521-530，1989）。但是，此处报道的数据表明，在携带rpoH，groEL或dnaK突变的大肠杆菌菌株中，高水平CytA产生不需要20-kDa蛋白，所有这些都影响细胞的蛋白水解能力。 20 kDa蛋白还增加了大肠杆菌生产的CryIVD（另一种苏云金芽孢杆菌亚种israelensis晶体蛋白）和LacZX90（β-半乳糖苷酶的突变体）的数量。后一种现象可归因于LacZX90的半衰期增加，这表明20 kDa的蛋白质可以稳定该蛋白质。还在体外和T7 RNA聚合酶表达系统中检测了20 kDa蛋白的作用，并讨论了这些结果对蛋白水解时间和20 kDa蛋白活性的可能意义。最后，单一抗体对大肠杆菌提取物中的CytA和20-kDa蛋白进行免疫沉淀的能力提供了可能与20-kDa蛋白作用机理有关的蛋白-蛋白相互作用的证据。

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