表达牛流行热病毒G基因的重组牛传染性鼻气管炎病毒的构建

Construction of recomtinant infectious bovine rhinotracheitis virus expressing bovine ephemeral fever virus glycoprotein g

病毒活载体疫苗兼有常规活疫苗和灭活疫苗的优点 ,是当今最有发展前景的疫苗研究领域之一。试验将已构建的表达牛流行热病毒 (BEFV)结构糖蛋白 G基因的牛传染性鼻气管炎病毒 (IBRV )转移载体 pd TK - L ac Z-G,通过脂质体方法转染由牛传染性鼻气管炎病毒 Bartha毒株感染的 MDBK细胞 ,筛选出形成蓝色蚀斑的重组病毒 ,并对其进行蚀斑纯化。通过 PCR方法鉴定该重组病毒证明基因组中含有完整的牛流行热病毒 G蛋白基因 ,实验为开发 BEFV/ IBRV二联基因工程疫苗奠定了基础。

The transmembrane glycoprotein G is the major neutralizing and protective antigen of bovine ephemeral fever rhabdovirus (BEFV).The transfer plasmid pdTK-LacZ-G with BEFV G gene and LzcZ reporter gene was used to transfect MDBK cell infected with IBRV using lipofectin.The recombintant virus was plaque-purified in the presence of X-gal for five rounds,and dentified by PCR method.This study can provide a basis for developing a genetically engineered vaccine candidate against BEFV/IBRV.