棉铃虫氨肽酶N基因片段克隆、表达和内源蛋白检测

Cloning,expression of aminopeptidase N(APN) from Helicoverpa armigeramidgut and identification of endogenous APN

氨肽酶N(APN)是苏云金芽孢杆菌杀虫毒素Cry在昆虫中肠中的一个重要受体。研究氨肽酶N在昆虫中肠中的分布特征对于阐明Cry毒素的杀虫机理和昆虫对Cry毒素的抗性机理具有重要的意义。通过RT-PCR的方法从棉铃虫中肠上皮细胞中克隆得到氨肽酶N的基因片段APN1551,并诱导表达纯化得到其重组蛋白APN517。以此蛋白为抗原,制备其抗血清。用该抗血清能检测到棉铃虫中肠上皮细胞中的APN蛋白。为研究Cry毒素的作用机理奠定基础。

Aminopeptidase N（APN） is one of important receptors in the insect midgut that Cry toxin from Bacillus thuringiensis（Bt） binds.To illuminate the mechanism of Cry toxin insecticidal action and inscet resistance to Cry toxin,it is necessary to study the distribution of APN in the insect midgut.In this paper,the APN gene fragment APN1551 by RT-PCR from Helicoverpa armigera midgut was cloned.This gene fragment was inserted into pGEX-4T-1 and the recombinant protein APN517 was induced and purified.Then the antiserum of APN517 was prepared.As the first antibody,this antiserum could recognize the endogenous APN protein in the Helicoverpa armigeramidgut.