摘要
将禽流感病毒H5N1亚型具有免疫原性的血凝素(HA)基因和神经氨酸酶(NA)基因,克隆于表达质粒Tmd-CMV-IRES-EGFP中,使其位于含细胞巨化病毒(CMV)早期启动子的调控下,并与增强型绿色荧光蛋白基因(EGFP)串联,将上述串联基因插入到火鸡疱疹病毒(HVT)基因的非必需区(US10)中,从而构建了带标记的重组转移质粒。将该转移载体质粒感染HVT的鸡胚成纤维细胞(CEF),利用脂质体共同转染CEF,待病毒蚀斑出现后,利用蚀斑筛选带有绿色荧光的重组病毒蚀斑,数轮蚀斑纯化,经荧光和PCR初步鉴定获得了重组火鸡疱疹病毒。
Haemagglutinin （HA） and neuraminidase （NA） genes of AIV were cloned in Tmd-CMV-IRES-EGFP. CMV-HA-IRES-NA-EGFP were inserted into the USIO gene region of herpesvirus of turkey（HVT）.Then the transfer vector plasmid and HVT DNA were cotransferred onto CEF cells. When virus plaque appeared three days later,the green fluorescent plaque was seen and picked. The recombinant turkey herpesvirus virus was purified by several cycles of plaque screening. By green fluorescence and PCR,the recombinant HVT virus was identified.
出处
《中国动物检疫》
CAS
北大核心
2008年第4期20-22,共3页
China Animal Health Inspection
基金
863项目(家禽重要病毒病基因工程疫苗研究和创新。课题编号:2006AA10A205)
