| 针对NSP4基因的shRNA抑制牛轮状病毒的体外增殖 |
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| 引用本文： | 陈方园,王洪梅,杨少华,武建明,高运东,刘晓,杨宏军,王长法,仲跻峰,王立群,何洪彬. 针对NSP4基因的shRNA抑制牛轮状病毒的体外增殖[J]. 病毒学报, 2010, 26(3) |
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| 作者姓名： | 陈方园 王洪梅 杨少华 武建明 高运东 刘晓 杨宏军 王长法 仲跻峰 王立群 何洪彬 |
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| 作者单位： | 东北农业大学生命科学学院,哈尔滨,150030;山东省农业科学院奶牛研究中心,济南,250100;山东省农业科学院奶牛研究中心,济南,250100;东北农业大学生命科学学院,哈尔滨,150030 |
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| 基金项目： | 现代农业产业(奶牛)技术体系岗位科学家(nycytx-0107); 国家转基因重大专项(2009ZX08007-006B); 山东省自然科学基金(Y2008D20),山东省自然基金(ZR2009DQ016); 山东农科院重大成果培育基金(2006YCG012); 山东省农业重大应用技术创新课题(何洪彬); 山东省科技攻关课题(2009GG20002032) |
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| 摘 要： | 本研究根据牛轮状病毒(Bovine rotavirus,BRV)的NSP4基因序列,设计并构建了shRNA重组慢病毒载体RNAi-H1-89,利用脂质体介导法,将RNAi-H1-89与辅助质粒共转染293细胞,包装shRNA重组慢病毒;重组慢病毒感染MA104细胞24h后继续接种BRV,以针对LacZ基因的shRNA重组慢病毒为对照,48h后与LacZshR-NA对照比较,RNAi-H1-89明显抑制MA104细胞病变;RT-PCR检测结果表明,针对NSP4基因的shRNA可特异性抑制牛轮状病毒NSP4基因表达;病毒滴度测定表明,在转染50%、75%、95%RNAi-H1-89质粒的细胞培养上清液中,病毒滴度分别是对照组的50%、20%和10%。上述实验结果表明,针对NSP4基因的shRNA能高效地沉默NSP4基因,并且能阻断牛轮状病毒增殖。
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| 关 键 词： | BRV shRNA NSP4基因 |
shRNA against NSP4 Gene Inhibits the Proliferation of Bovine Rotavirus in vitro |
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| CHEN Fang-yuan,,WANG Hong-mei,YANG Shao-hua,WU Jian-ming,GAO Yun-dong,LIU Xiao,YANG Hong-jun,WANG Chang-fa,ZHONG Ji-feng,WANG Li-qun,HE Hong-bin. shRNA against NSP4 Gene Inhibits the Proliferation of Bovine Rotavirus in vitro[J]. Chinese journal of virology, 2010, 26(3) |
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| Authors: | CHEN Fang-yuan WANG Hong-mei YANG Shao-hua WU Jian-ming GAO Yun-dong LIU Xiao YANG Hong-jun WANG Chang-fa ZHONG Ji-feng WANG Li-qun HE Hong-bin |
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| Affiliation: | CHEN Fang-yuan1,2,WANG Hong-mei2,YANG Shao-hua2,WU Jian-ming2,GAO Yun-dong2,LIU Xiao2,YANG Hong-jun2,WANG Chang-fa2,ZHONG Ji-feng2,WANG Li-qun1,HE Hong-bin2 (1.Life College of Northeast Agricultural University,Harbin 150030,China,2. Shandong Province Academy of Agricultural Sciences Dairy Cattle Research Center,Jinan 250100,China) |
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| Abstract: | Based on the NSP4 sequence of bovine rotavirus (BRV),the shRNA was designed and synthesized,and a shRNA recombinant lenti-virus vector RNAi-H1-89 was constructed. The recombinant RNAi-H1-89 Lenti-virus was packaged by transfecting the 293T cell with the recombinant vector RNAi-H1-89 and two helper plasmids using lipofectamine,and then used to infect MA104 cells. The MA104 cells were further infected with BRV strain G6 24h post-infection,with the LacZ shRNA recombinant lenti-virus as control. Thirty -six hou... |
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| Keywords: | BRV shRNA NSP4 gene |
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