狂犬/丙型肝炎嵌合病毒的构建和拯救

目的 以狂犬病病毒为载体,构建表达丙型肝炎病毒包膜糖蛋白E1E2的狂犬/丙型肝炎嵌合病毒,为发展新型丙肝载体疫苗奠定基础.方法 在狂犬病病毒反向遗传系统CTN-GFP的基础上,通过传统分子克隆方法,将HCV E1E2基因分别克隆人复制型和复制缺陷型狂犬病病毒载体,构建狂犬/丙肝嵌合病毒CTN-HCV E1E2和CTNΔG-HCV E1E2.结果 免疫荧光（DFA）和反转录聚合酶链式反应（RT-PCR）结果 显示嵌合病毒拯救成功,嵌合病毒能够再次感染正常细胞并且能够在mRNA水平检测到HCV E1E2基因的表达.结论 本研究成功构建了表达丙型肝炎病毒包膜糖蛋白E1E2的狂犬/丙肝嵌合病毒,提示以狂犬病病毒为载体发展新型丙肝载体疫苗在理论和技术上都是可行的.

Construction and recovery of chimeric rabies virus expressing envelop proteins E1E2 of hepatitis C

Objective Construction and recovery of chimeric rabies virus expressing HCV envelop proteins E1E2. Methods On the basis of the previously established reverse genetic system CTN-CFP, HCV E1 E2 genes were cloned to both replication competent and replication constrained viral vectors based on CTN181 strain and the chimeric viruses CTN-HCV E1E2 and CTNAG-HCV E1E2 were recovered. Results The result demonstrated that both the chimeric viruses were rescued successfully, had the ability to reinfect normal sensitive cell lines and express HCV E1E2 genes detected in the level of mRNA. Conclusion The establishment of chimeric RVs expressing HCV E1E2 genes provides the evidence that it is feasible to develop novel HCV vaccines based on viral vectors in theory and in practice.