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Efficient expression of the mosquito larvicidal binary toxin gene from Bacillus sphaericus in Escherichia coli

机译:球形芽孢杆菌的蚊幼虫二元毒素基因在大肠杆菌中的高效表达

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The binary toxin gene encoding BinA (42 kDa) and BinB (51 kDa) from Bacillus sphaericus strain 2297 was cloned and expressed in E. coli. Low expression level was found when both proteins were expressed from a single operon. High expression was observed when the gene encoding an individual protein was placed downstream of the T7 promoter. The expression level of BinB was not different when expressed alone (non-fusion) or as a fusion form with T7 peptide (T7-BinB). Both forms of BinB were equally stable. Unlike BinB, the non-fusion form of BinA was less stable than T7-BinA. The mosquito larvicidal test showed that BinA or BinB alone was not toxic to mosquito larvae, but high toxicity was found when both BinA and BinB were applied. The results suggest that a short peptide of T7 linked to the N-terminus of either BinA or BinB does not affect their toxicity, but may make the toxin, especially BinA, more stable.
机译:克隆了来自球形芽孢杆菌菌株2297的编码BinA(42 kDa)和BinB(51 kDa)的二元毒素基因,并在大肠杆菌中表达。当两种蛋白质都由一个操纵子表达时,发现表达水平较低。当将编码单个蛋白质的基因置于T7启动子的下游时,观察到高表达。当单独表达(非融合)或与T7肽(T7-BinB)融合时,BinB的表达水平没有差异。 BinB的两种形式均稳定。与BinB不同,BinA的非融合形式不如T7-BinA稳定。蚊虫幼虫试验表明,单独的BinA或BinB对蚊虫幼虫无毒,但同时使用BinA和BinB时发现高毒性。结果表明,与BinA或BinB的N端连接的T7短肽不会影响其毒性,但可能会使毒素(尤其是BinA)更稳定。

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