Identification of a second cytotoxic protein produced by Bacillus thuringiensis A1470
Identification of a second cytotoxic protein produced by Bacillus thuringiensis A1470
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DOI：
10.1007/s10529-013-1275-6
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发表时间：
2013-11-01
影响因子：
2.7
通讯作者：
Mizuki, Eiichi
中科院分区：
文献类型：
--
作者：
Okumura, Shiro;Ishikawa, Tomoyuki;Mizuki, Eiichi
苏云金芽孢杆菌A1470产生多种具有相似分子量（类似于30 kDa）的蛋白质，对人类细胞系具有细胞毒性。其中一种是以前鉴定的parasporin-4，是一种β孔形成毒素。第二种细胞毒性蛋白的N-末端序列与由B产生的parasporin-2的部分序列相同。苏云金杆菌A1547。通过使用副孢子蛋白-2的引物对来自A1470的总质粒DNA进行PCR以扩增基因，然后克隆该基因。克隆的基因与A1547 parasporin-2相差8 bp，预测的蛋白质相差4个氨基酸。该基因在大肠杆菌中表达，重组蛋白对四种人细胞系（MOLT-4，Jurkat，HeLa和HepG 2）的细胞毒活性与A1547 parasporin-2相似。然后，我们证实了A1470菌株同时产生parasporin-2和parasporin-4。
Bacillus thuringiensis A1470 produces multiple proteins with similar molecular masses (similar to 30 kDa) with cytotoxicity against human cell lines. One that was previously identified, parasporin-4, is a beta-pore-forming toxin. The N-terminal sequence of a second cytotoxic protein was identical to a partial sequence of parasporin-2 produced by B. thuringiensis A1547. PCR was performed on total plasmid DNA from A1470 by using primers for parasporin-2 to amplify a gene which was then cloned. The cloned gene differed from A1547 parasporin-2 by 8 bp and the predicted protein differed by four amino acids. The gene was expressed in Escherichia coli, and the cytotoxic activities of the recombinant protein against four human cell lines (MOLT-4, Jurkat, HeLa, and HepG2) were similar to those of A1547 parasporin-2. We then confirmed that the A1470 strain simultaneously produces parasporin-2 and parasporin-4.