埃可病毒30型重组蛋白vp1表达及其在病原检测中的应用  

expression of recombinant protein vp1and its application in developing enzyme-linked immunoassay for echovirus 30

作　　者：朱坚胜[1] 阮仙利[1] 汪志军[1] 周林福[2] 王昕昕[1] 王丽珍[1] 陈华忠[1] 王赓歌[1] 

机构地区：[1]浙江省台州医院感染科,317000 [2]浙江大学医学院

出　　处：《中华临床感染病杂志》2012年第2期81-84,84,97,117,共4页Chinese Journal of Clinical Infectious Diseases

基　　金：浙江省科技计划重点课题资助

摘　　要：目的 表达埃可病毒30型vp1重组蛋白,建立埃可病毒特异性抗体的检测方法.方法 采用反转录聚合酶链反应(rt-pcr)方法获得埃可病毒vp1cdna,克隆至原核表达载体pet44a中表达,并用纯化的重组蛋白检测血清中埃可病毒抗体.结果 表达的重组蛋白vp1相对分子质量约为95 000,经western blot和酶联免疫吸附试验证实其具有良好的抗原性.结论 基因工程重组融合蛋白vp1可以作为诊断用抗原.objective to express vp1recombinant protein of echovirus 30 (echo30) in e.coli bl21(de3) and to develop an enzyme-linked immunoassay (eia) based on the recombinant antigen for detecting antibodies to echo30.methods the target vp1gene was amplified by reverse transcription-polymerase chain reaction (rt-pcr).the pcr products of the gene were inserted into pet44a vector,and then expressed in e.coli bl21( de3 ).the purified recombinant protein was used for the development of eia.results the molecular weight of the recombinant protein was 95 000,and the antigenicity of which was identified by western blot and eia.conclusion the recombinant protein vp1has been successfully expressed and purified,which can be used as diagnostic antigen.

关 键 词：重组蛋白质类 酶联免疫吸附测定 埃可病毒 

分 类 号：R4[医药卫生—临床医学] R51