利用反向遗传技术构建表达绿色荧光蛋白的H9N2禽流感重组病毒

GENERATION OF A RECOMBINANT H9N2 INFLUENZA VIRUS EXPRESSING GFP BY REVERSE GENETIC MANIPULATION

H9N2亚型禽流感病毒普遍存在于我国大部分地区,宿主范围较广,是世界卫生组织发布的最具大流行潜力的病毒之一。为了利用H9N2亚型禽流感病毒表达绿色荧光蛋白(green fl uorescent protein,GFP),通过反向遗传操作技术将A/Chicken/Shanghai/2093/2009(H9N2)(SH2093)毒株的NS2基因连接到PB1基因下游,同时将GFP基因连接在NS1基因之后,拯救并获得了1株表达GFP的重组病毒SH2093-GFP。虽然SH2093-GFP在MDCK细胞上的增殖能力低于野生毒株SH2093,但该病毒可以成功表达GFP,为进一步研究H9N2亚型禽流感病毒作为外源基因表达载体等提供重要依据。

The H9N2 Avian influenza virus （AIV） circulating in most parts of China with a wide host range, has been on the list of World Health Organization for the greatest pandemic potential. To obtain a HgN2 AIV expressing enhanced green-fluorescent protein （GFP）, reverse genetics system was used to manipulate the AIV strain A/Chicken/Shanghai/2093/2009 （H9N2） （SH2093） in order to develop the recombinant virus （SH2093-GFP）. Rearrangement of the influenza genome was accomplished by inserting the NS2 gene downstream of the PB 1 gene to express NEP protein and G FP gene downstream of a full-length NS1 gene. The recombinant virus SH2093-GFP expressed GFP on MDCK cells successfully though its replication was not efficient as compared with its parental virus SH2093, suggesting that the H9N2 AIV might be a promising vector candidate for expression of other exogenic proteins.