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

摘要:

本研究在完成FMDV O/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, 48 h后收获培养液接种2~3 d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。

Abstract:

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 carried 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.