State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100193, People's Republic of China.
Biotechnol Lett. 2011 Mar;33(3):531-7. doi: 10.1007/s10529-010-0452-0. Epub 2010 Nov 3.
cry1Ah1, a novel holo-type gene cloned from Bacillus thuringiensis strain BT-8, encoded a protein exhibiting strong insecticidal activity against lepidopteran insects. To identify the minimal active fragment of the Cry1Ah toxin, 9 pairs of primers were designed to generate different PCR products. Seven PCR products were amplified by different primers using the cry1Ah1 gene as a template and cloned into a pET-21b vector. These positive clones were separately transformed into Escherichia coli. Insecticidal activity against 2nd-instar larvae of Plutella xylostella was performed using the leaf-dip bioassay: the minimal active fragment of the Cry1Ah toxin was located between amino acid residues 50I and 639E.
cry1Ah1 是从苏云金芽孢杆菌菌株 BT-8 中克隆出的一种新型全孢型基因，编码的蛋白对鳞翅目昆虫具有很强的杀虫活性。为了鉴定 Cry1Ah 毒素的最小活性片段，设计了 9 对引物，以产生不同的 PCR 产物。使用 cry1Ah1 基因作为模板，用不同的引物扩增了 7 个 PCR 产物，并将其克隆到 pET-21b 载体中。将这些阳性克隆分别转化到大肠杆菌中。采用叶浸生物测定法测定对小菜蛾 2 龄幼虫的杀虫活性：Cry1Ah 毒素的最小活性片段位于 50I 至 639E 氨基酸残基之间。
