Belfiore C J, Vadlamudi R K, Osman Y A, Bulla L A
Department of Molecular Biology, University of Wyoming, Laramie 82071.
Biochem Biophys Res Commun. 1994 Apr 15;200(1):359-64. doi: 10.1006/bbrc.1994.1456.
Biopesticides based on the bacterium Bacillus thuringiensis have attracted wide attention as safe alternatives to chemical insecticides. In this paper, we report, for the first time, the identification of a single binding protein from a coleopteran insect, Tenebrio molitor, that is specific for the cryIII toxin of B. thuringiensis. The protein appeared as a single band of 144 kDa on radioligand and immunoblots of total proteins extracted from brush border membrane vesicles of the midgut of T. molitor. Radiolabelled cryIIIA toxin bound to the protein with a Kd value of 17.5 nM and could be specifically blocked by unlabelled toxin but not by toxins from other subspecies of B. thuringiensis. This study lays the groundwork to clone the cryIIIA toxin binding protein and to determine the molecular mechanism(s) of toxin action.
基于苏云金芽孢杆菌的生物农药作为化学杀虫剂的安全替代品已引起广泛关注。在本文中,我们首次报道了从鞘翅目昆虫黄粉虫中鉴定出一种对苏云金芽孢杆菌cryIII毒素具有特异性的单一结合蛋白。该蛋白在从黄粉虫中肠刷状缘膜囊泡提取的总蛋白的放射性配体和免疫印迹上呈现为一条144 kDa的单带。放射性标记的cryIIIA毒素以17.5 nM的Kd值与该蛋白结合,并且可以被未标记的毒素特异性阻断,但不能被苏云金芽孢杆菌其他亚种的毒素阻断。这项研究为克隆cryIIIA毒素结合蛋白和确定毒素作用的分子机制奠定了基础。