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口蹄疫病毒O/HN/93疫苗株的拯救及病毒活性鉴定

DOI: 10.7668/hbnxb.2010.03.008, PP. 32-37

Keywords: 口蹄疫病毒,疫苗株,感染性cDNA克隆,病毒拯救

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Abstract:

利用定点突变方法,构建含有预期突变的口蹄疫病毒O/HN/93株全长cDNA克隆,将全长cDNA的重组质粒线化后,与表达T7RNA聚合酶的真核质粒pcDNAT7P共转染BHK-21细胞,转染细胞盲传至第3代50h后出现典型致细胞病变效应(CPE),第4代10h出现典型的CPE。对收获的病毒分别用RT-PCR、分子标签测序、间接免疫荧光和电镜观察等进行鉴定,均证实成功拯救到了O/HN/93株FMDV。成功获得O/HN/93株的感染性分子克隆,为进一步研究抗原谱广、免疫原性好的候选疫苗株奠定了骨架基础。

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