H7亚型禽流感病毒HA1基因的克隆与原核表达  

Cloning and Prokaryotic Expression of HA1 Gene of H7 Avian Influenza Virus

作　　者：庞平[1] 唐懿[1] 雷霆[1] 武山[1] 李强[1] 陈晨[1] 曹红[1] 陈福勇[1] 

机构地区：[1]中国农业大学动物医学院,北京100094

出　　处：《中国家禽》2006年第23期10-13,共4页China Poultry

基　　金：国家科技攻关计划课题(2004BA519A18)

摘　　要：利用PCR技术亚克隆了H7亚型禽流感病毒的HA1基因,并将PCR产物连接到克隆载体pMD 18-T Simple vector,转化大肠杆菌。测序结果表明所克隆的片断大小为1035bp。将HA1基因克隆至原核表达载体pGEX-6P-1,经酶切和PCR鉴定,证明成功构建了重组表达载体pGEX-HA1。将构建好的融合表达载体在IPTG的诱导下在大肠杆菌中得到了表达。融合蛋白GST-HA1的分子量为63ku。Western-Blot和ELISA鉴定结果表明,融合蛋白与H7亚型禽流感阳性血清发生特异性反应,而与H5、H9亚型抗血清不发生反应。In this study,HA1 gene was subcloned by PCR,ligated into pMD18-T simple vector and then transformed E.coli. C, ene sequencing analysis showed that the cloned HA1 was 1 035 bp in length. The HA1 gene was inserted into the expression plasmid pGEX-6P-1. The recombinant fusion protein was expressed in E.coli BL21 induced by IPTG in the form of inclusion bedies, The molecular weights of GST-HA1 is 63 ku as demonstrated by western-blotting using AIV antibody against H7 subtype. The result of ELISA developed with this recombinant protein showed that the recombinant protein can only react with antisera of H7 subtype.

关 键 词：禽流感病毒 HA1基因 基因克隆 表达 抗原性 

分 类 号：S855.3[农业科学—临床兽医学]