口蹄疫病毒O/QYYS/s/06株感染性克隆的构建  

Construction of an infectious cDNA clone derived from foot-and-mouth disease virus O/QYYS/s/06

作　　者：卢受昇[1,2] 赵启祖[3] 刘湘涛[4] 孙彦伟 任涛[1] 张桂红[1] 亓文宝[1] 查云峰 孔令辰 张翰 樊惠英[1] 廖明[1] 

机构地区：[1]华南农业大学兽医学院,广州510642 [2]广东省动物防疫监督总所,广州510230 [3]中国兽医药品监察所,北京100081 [4]中国农业科学院兰州兽医研究所,兰州730046

出　　处：《生物工程学报》2009年第7期982-986,共5页Chinese Journal of Biotechnology

基　　金：国家自然科学基金(No.30800826)资助~~

摘　　要：本研究在完成FMDVO/QYYS/s/06株全基因组序列测定的基础上,分3段对全基因组进行克隆,其后将各片段克隆到载体P43中,从而获得携带O/QYYS/s/06株基因组全长cDNA的重组质粒P43C。将重组质粒P43C与表达RNA聚合酶的质粒T7共转染BHK-21细胞,48h后收获培养液接种2~3d乳鼠,取经乳鼠传代后的第4代病毒液,经反向间接血凝、中和试验和测序等方法证明拯救的病毒为O型FMDV。以上结果表明,O/QYYS/s/06株全长cDNA分子克隆的构建成功。After sequencing, we amplified and cloned foot-and-mouth disease virus （FMDV） O/QYYS/s/06 whole genome by three fragments. These three fragments were cloned into vector P43 one by one to construct recombinant plasmid P43C, which carded the full-length cDNA of FMDV O/QYYS/s/06. Then, plasmid P43C and plasmid T7 expressing T7 RNA polymerase were co-transfected into BHK-21 cells. After 48 h, we harvested the culture broth from transfected BHK-21 cells and inoculated into 2-3 day-old sucking mice. After four generation passage, the virus harvested from sucking mice was confirmed to be type O FMDV by the indirect hemagglutination test, sucking mice＇s neutralization test and sequencing. The results showed that we have successfully constructed the full-length cDNA clone of FMDV O/QYYS/s/06 strain.

关 键 词：口蹄疫病毒 O/QYYS/s/06株 全长CDNA 

分 类 号：S852.65[农业科学—基础兽医学]