作　　者： ; ; ; ; ; ; ; ;

机构地区： 中国农业大学动物医学院

出　　处： 《中国家禽》 2006年第23期10-13,共4页

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 基因 基因克隆 表达 抗原性

领　　域： [农业科学] [农业科学] [农业科学]