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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
引用本文:刘振江,鲁会军,李国江,刘昊,靖杰,刘燕瑜,岳云强,郭欢欢,凡敏,秦艳青,金宁一.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[J].中国兽医学报,2012,32(8):1094-1097,1100.
作者姓名:刘振江  鲁会军  李国江  刘昊  靖杰  刘燕瑜  岳云强  郭欢欢  凡敏  秦艳青  金宁一
作者单位:1. 吉林农业大学动物科技学院,吉林长春130118/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
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. 吉林农业大学动物科技学院,吉林长春130118/吉林农业科技学院,吉林吉林市132101
4. 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/吉林大学畜牧兽医学院,吉林长春130062
基金项目: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。

关 键 词:猪源新城疫病毒  F和HN基因  T7  RNAP  细胞融合

Expression of F and HN genes of swine Newcastle disease virus and its fusion effect on BHK-21 cell
LIU Zhen-jiang,LU Hui-jun,LI Guo-jiang,LIU Hao,JING Jie,LIU Yan-yu,YUE Yun-qiang,GUO Huan-huan,FAN Min,QIN Yan-qing,JIN Ning-yi.Expression of F and HN genes of swine Newcastle disease virus and its fusion effect on BHK-21 cell[J].Chinese Journal of Veterinary Science,2012,32(8):1094-1097,1100.
Authors:LIU Zhen-jiang  LU Hui-jun  LI Guo-jiang  LIU Hao  JING Jie  LIU Yan-yu  YUE Yun-qiang  GUO Huan-huan  FAN Min  QIN Yan-qing  JIN Ning-yi
Institution:1.College of Animal Science,Jilin Agricultural University,Changchun 130118,China;2.Key Laboratory of Jilin Provice for Zoonosis Prevention and Control,Institute of Military Veterinary,Academy of Military Medical Science,Changchun 130122,China;3.Jilin Agricultural and Technology College,Jilin City,Jilin 132101,China;4.College of Animal Science and Veterinary Medicine,Jilin University,Changchun 130062,China)
Abstract: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 according 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-T7-EGFP were co-transfected into BHK-21 cells.16 h after transfection,the transfected cells showed significant cell fusion by Giemsa staining.This study laid a foundation for study on F and HN gene of swine Newcastle disease virus.
Keywords:swine NDV  F and HN gene  T7 RNAP  cell fusion
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