狂犬病毒aG株糖蛋白在pET原核系统中的表达及纯化  

The expression and purification of glycoprotein of rabies virus aG strains in pET prokaryotic system

作　　者：何珺 严家新[1] 李承平[1] 朱家鸿[1] 

机构地区：[1]卫生部武汉生物制品研究所

出　　处：《微生物学免疫学进展》1999年第3期1-6,共6页Progress In Microbiology and Immunology

摘　　要：在狂犬病的临床诊断和基础研究中都迫切需要大量纯度高且价廉的狂犬病毒糖蛋白抗原。本文应用带有Ｈｉｓ６尾的ｐＥＴ原核表达系统对狂犬病毒（ＲＶ）ａＧ株的糖蛋白（ＧＰ）进行表达和纯化。构建的融合表达载体ｐＥＴ－ａＧ１和ｐＥＴ－ａＧ２（－５７ｂｐ）分别含有ＲＶａＧ株ＧＰ基因的全序列及删除了为ＧＰ信号肽编码的５８个碱基的序列。用ＳＤＳ聚丙烯酰胺凝胶电泳、免疫印迹、间接ＥＬＩＳＡ检测都证明表达产物为ＲＶＧＰ，且位于菌体中的包涵体内。经固定化金属螯合层析（ＩＭＡＣ）提纯，ｐＥＴ－ａＧ１表达产物有较高的特异性和纯度，可用作测定ＲＶＧＰ抗体的免疫诊断试剂。In basic research and clinical diagnosis of rabies virus,there is an urgent need for rabies virus(RV) glycoprotein(GP) with high purity and low cost.The GP of RV aG strain were expressed and purified by pET expression systems in E.coli.Both the plasmids pET aG1 and pET aG2(-57bp) constructed contained the cDNA of RV GP gene,but the latter had a 58 bp deletion which encoded for the signal peptids.It was demonstrated by SDS-PAGE,Westen-blot and ELISA that the expression products were RV GP and they appeared in the inclusion body of host cells.The expression product of pET aG1 purified by IMAC possessed high purity and specificity and it could be used in diagnostic ELISA for RV GP.

关 键 词：狂犬病毒 糖蛋白 融合表达系统 金属螯合层析 

分 类 号：R373.9[医药卫生—病原生物学]