摘要： 以本实验室保存的REV毒株为模板，采用RT-PCR技术扩增了REV env部分基因，长为939 bp，并进行亚克隆到pMD18-T质粒载体上，连接、转化、鉴定后得到阳性重组质粒pET-env，将目的片段进行序列分析，结果表明，该REV的env基因与已发表的SNV株、中国HA9901株以及A株同源性均达94%以上，推导的氨基酸同源性为94%以上。将该基因片段与原核表达载体pET-32a重组，并将重组质粒转化至宿主菌BL21中，用IPTG诱导表达，对表达的蛋白用SDS-PAGE电泳，免疫印迹和薄层凝胶扫描分析，表达产物与理论推理一致；免疫印迹结果证明，表达的env蛋白可被REV阳性血清所识别。

关键词: 禽网状内皮组织增殖病病毒, env基因, 克隆, 原核表达

Abstract: The viral sub-genome mRNA of Avian Reticuloendotheliosis Virus was amplified with RT-PCR. A DNA fragment was amplified which contains a part of env gene, and the size of the DNA fragment about 939 bp. The PCR products were purified and then cloned into plasmid pMD18-T, the recombinant plasmid were designated Pmd-env and analyzed by endonecleoase digestion from proper inserts. The sequence analysis of the insert fragment in the recombinant pMD-env indicated that env shared more than 94% with SNV strain, HA9901 strain, and A strain. The amino acid sequence homologywas above 94%. The env gene was subcloned into a prokaryotic expressing vector, pET-32a. Molecular cloning of env gene from REV provides a basis for the studies on protein expressing and molecular characteristic, and the constructed recombinant pET-env can be used in further protein expressing.

Key words: REV, envgene, Cloning, Prokaryotic expressing

中图分类号: 

• S432.4+1