猪源新城疫病毒F和HN基因的表达及其对BHK-21细胞膜的融合作用  

Expression of F and HN genes of swine Newcastle disease virus and its fusion effect on BHK-21 cell

作　　者：刘振江[1,2] 鲁会军[2] 李国江[1,3] 刘昊[2,4] 靖杰[2,4] 刘燕瑜[2,4] 岳云强[1,2] 郭欢欢[1,2] 凡敏[1,2] 秦艳青[1,2] 金宁一[2] 

机构地区：[1]吉林农业大学动物科技学院,吉林长春130118 [2]军事医学科学院军事兽医研究所吉林省人兽共患病预防与控制重点实验室,吉林长春130122 [3]吉林农业科技学院,吉林吉林市132101 [4]吉林大学畜牧兽医学院,吉林长春130062

出　　处：《中国兽医学报》2012年第8期1094-1097,1100,共5页Chinese Journal of Veterinary Science

基　　金：国家"973"计划资助项目(2011CB504702);国家自然科学基金资助项目(30800828)

摘　　要：为了分析猪源新城疫病毒F和HN基因对BHK-21细胞膜的融合作用,采用RT-PCR方法从猪源新城疫病毒JL01株中扩增获得了F蛋白基因,并根据GenBank上登录的猪源新城疫病毒HN基因序列,合成了HN基因,分别将F和HN基因克隆到pKS载体上。又从大肠杆菌BL21(DE3)中扩增得到T7RNA聚合酶基因,将报告基因EGFP与T7RNA聚合酶基因一起定向克隆到痘苗病毒转移载体pSTK上。再将pKS-F、pKS-HN与pSTK-T7-EGFP共转染BHK-21细胞,转染后16h,用Giemsa染液进行染色,可见明显的细胞融合现象。该研究证明猪源新城疫病毒JL01株的F和HN蛋白共同作用于BHK-21细胞使细胞膜融合。To investigate the effect of expressing F and HN genes of swine Newcastle disease virus JL01 strain on BHK-21 cell fusion,we got F gene from swine Newcastle disease virus by RT-PCR, and synthesized HN gene ac- cording to swine Newcastle disease virus HN gene sequence from the GenBank. And then the F and HN genes were inserted into the pKS vector, respectively. T7 RNAP gene was amplified from E. coli BL21 （DE3）, and along with the EGFP gene was inserted into pSTK vector to construct recombinant plasmid pSTK-T7-EGFP, pKS-F, pKS-HN and pSTK-TT-EGFP were co-transfected into BHK-21 cells. 16 h after transfection,the transfected cells showed sig- nificant cell fusion by Giemsa staining. This study laid a foundation for study on F and HN gene of swine Newcastle disease virus.

关 键 词：猪源新城疫病毒 F和HN基因 T7 RNAP 细胞融合 

分 类 号：S852.65[农业科学—基础兽医学]