马铃薯Y病毒HC-Pro基因的克隆、序列分析以及原核表达  

Cloning and Sequencing of Helper Component Proteinase Gene of Potato Virus Y and its Expression in Escherishia coli

作　　者：郭明[1] 鲁瑞芳[1] 王海云[1] 彭学贤[1] 魏宁生[2] 莽克强[1] 

机构地区：[1]中国科学院微生物所植物生物技术实验室,北京100080 [2]西北农业大学植保系,陕西扬凌712100

出　　处：《农业生物技术学报》2000年第3期249-252,共4页Journal of Agricultural Biotechnology

基　　金：中国科学院微生物所植物生物技术开放实验室资助项目

摘　　要：本研究以马铃薯Y病毒中国株系(PVY-C) RNA 为模板，通过RT-PCR 克隆了PVY-C HC-Pro基因,并对其进行了序列分析。测序结果表明PVY-C HC-Pro基因全长为1 368 nt，编码一个含456个氨基酸的蛋白。它与PVY其它株系的同源性高于与马铃薯Y病毒属其它种病毒的同源性,与所报道的PVYn株系的碱基及氨基酸序列同源性均为96 %，推测PVY-C可能是PVYn株系或它的一个近缘株系。SDS-PAGE表明在大肠杆菌中表达了PVY-C HC-Pro, Western blot 证明将Ni2+-NTA 树脂纯化获得的特异表达产物制备了HC-Pro的抗血清。The full-length helper component proteinase(HC-Pro)gene of p otato virus Y of Chinese strain(PVY-C)was cloned and its complete nucl eotide sequence was determined. The results showed that it is composed of 1 368 nt, which encodes a protein of 456 amino acids. It shares hi gher homology with PVY other strains than with other viruses of potyvi rus, and the highest homology of 96 % with PVYn strain in both nucleot ide and putative amino acid sequences. The complete cDNA of PVY-C HC-P ro was then cloned into expression vector pBV221, pQE30 and pTrcHisA, respectively. SDS-PAGE showed that it was successfully expressed in E. coli, and Western blot analysis demonstrated that the antiserum of HC- Pro was raised against the specific expression product purified by usi ng Ni2+- NTA resin.

关 键 词：马铃薯Y病毒 HC-PRO基因 序列分析 

分 类 号：S432.41[农业科学—植物病理学] S435.32[农业科学—农业昆虫与害虫防治]