荧光蛋白标记表达H6亚型禽流感病毒血凝素基因

Expressing of HA Gene of H6 Subtype Avian Influenza Virus with Fluorescent Protein Marked

研究构建了H6N2亚型AIV的核酸疫苗表达载体,以绿色荧光蛋白基因作为报告基因,构建了HA与报告基因的融合基因,并克隆于pAVX1表达载体,再经脂质体转染BHK细胞进行体外表达试验。构建的表达载体pAVX1-H6A-GFP酶切鉴定已克隆上融合基因;体外表达试验中,脂质体转染24 h后出现微弱荧光,48 h后见较强的荧光表达,从而成功实现了用荧光蛋白标记表达了H6亚型禽流感病毒血凝素基因。

H6N2 subtype AIV DNA vaccine expression vector was constructed in this research,the vaccine was marked with green fluorescent protein. The HA-GFP fusion gene was cloned on plasmid pAVX1 expression vector,the vaccine was expressed in BHK cell by lipofection .The expression vector pAVX1-H6A-GFP succeeded to clone the fusion gene HA-GFP by enzymolysis identification. In vitro expression tests, the cells got weak fluorescence after lipofection 24 hours,and got strong fluorescence after lipofection 48 hours. This indicated the research succeeded to express HA gene of H6 subtype Avian influenza virus of HA gene with green fluorescent protein marked.