Milne R E, Pang A S, Kaplan H
Forest Pest Management Institute, Natural Resources Canada, Sault Ste Marie, Ontario, Canada.
Insect Biochem Mol Biol. 1995 Dec;25(10):1101-14. doi: 10.1016/0965-1748(95)00046-1.
A 75 kDa protein from spruce budworm (Choristoneura fumiferana) gut-juice has been isolated and shown to cause a specific precipitation of the delta-endotoxin from Bacillus thuringiensis subsp. sotto. This 75 kDa protein, separated by either column chromatography or SDS-PAGE, caused precipitation of the sotto toxin in both agarose diffusion gels and the PAGE gels. The precipitation event leads to limited proteolysis of the toxin and loss of larval toxicity. SDS-PAGE analysis of the precipitated toxin indicates that proteolysis of the toxin is not a prerequisite for precipitation. The protein responsible for precipitation, exhibits elastase-like activity and appears to be a complex which partially dissociates during boiling in SDS-PAGE sample buffer. Gut-juice from gypsy moth, forest tent caterpillar and white mark tussock moth also precipitated delta-endotoxin, but silkworm gut-juice gave a much weaker response. These results provide further evidence that, in the larval gut, differential processing of delta-endotoxin may play a role in the expression of activity towards various insect larvae.
从云杉芽卷叶蛾(Choristoneura fumiferana)肠道汁液中分离出一种75 kDa的蛋白质,该蛋白质可使苏云金芽孢杆菌 sotto亚种的δ-内毒素发生特异性沉淀。通过柱色谱法或SDS-PAGE分离得到的这种75 kDa蛋白质,在琼脂糖扩散凝胶和PAGE凝胶中均能使sotto毒素沉淀。沉淀过程导致毒素发生有限的蛋白水解,并丧失幼虫毒性。对沉淀毒素的SDS-PAGE分析表明,毒素的蛋白水解并非沉淀的先决条件。负责沉淀的蛋白质具有类弹性蛋白酶活性,似乎是一种复合物,在SDS-PAGE样品缓冲液中煮沸时会部分解离。舞毒蛾、森林天幕毛虫和白纹毒蛾的肠道汁液也能沉淀δ-内毒素,但家蚕肠道汁液的反应则弱得多。这些结果进一步证明,在幼虫肠道中,δ-内毒素的差异加工可能在对各种昆虫幼虫的活性表达中起作用。