Okumura Shiro, Saitoh Hiroyuki, Ishikawa Tomoyuki, Mizuki Eiichi, Inouye Kuniyo
Fukuoka Industrial Technology Centre, Kurume, Fukuoka 839-0861, Japan.
Biotechnol Annu Rev. 2008;14:225-52. doi: 10.1016/S1387-2656(08)00009-4.
In 1901, a unique bacterium was isolated as a pathogen of the sotto disease of the silkmoth larvae, and later in 1915, the organism was described as Bacillus thuringiensis. Since the discovery, this bacterium has widely attracted attention of not only insect pathologists but many other scientists who are interested in strong and specific insecticidal activity associated with inclusion bodies of B. thuringiensis. This has led to the recent worldwide development of B. thuringiensis-based microbial insecticides and insect-resistant transgenic plants, as well as the epoch-making discovery of parasporin, a cancer cell-specific cytotoxin. In the review, we introduce a detection study of interaction between inclusion proteins of B. thuringiensis and brush border membrane of insects using surface plasmon resonance-based biosensor, and then identification and cloning of parasporin-4, a latest cancer cell-killing protein produced by B. thuringiensis A1470 strain. Inclusion bodies of the parasporin-4 produced by recombinant Escherichia coli were solubilized and activated with a new method and purified by an anion-exchange chromatography. At last the characterization of the recombinant parasporin-4 was shown.
1901年，一种独特的细菌被分离出来，它是家蚕幼虫 sotto 病的病原体，后来在1915年，该微生物被描述为苏云金芽孢杆菌。自发现以来，这种细菌不仅引起了昆虫病理学家的广泛关注，也吸引了许多其他对苏云金芽孢杆菌包涵体相关的强大而特异的杀虫活性感兴趣的科学家。这导致了近年来基于苏云金芽孢杆菌的微生物杀虫剂和抗虫转基因植物在全球范围内的发展，以及芽孢杆菌蛋白（一种癌细胞特异性细胞毒素）这一划时代的发现。在这篇综述中，我们介绍了一项利用基于表面等离子体共振的生物传感器对苏云金芽孢杆菌包涵体蛋白与昆虫刷状缘膜之间相互作用的检测研究，然后对苏云金芽孢杆菌A1470菌株产生的最新癌细胞杀伤蛋白芽孢杆菌蛋白-4进行了鉴定和克隆。重组大肠杆菌产生的芽孢杆菌蛋白-4的包涵体用一种新方法进行溶解和活化，并用阴离子交换色谱法进行纯化。最后展示了重组芽孢杆菌蛋白-4的特性。
