【摘要】 设计并合成了4条O型口蹄疫病毒(FMDV)泛亚型代表毒株O/China/99基因组的引物,利用RT-PCR扩增各基因片段,酶切后连接到pOK12载体上,然后人工合成一段缺失PKs的5′UTR序列,利用两端的限制性内切酶位点,将该基因片段插入到上述载体中。酶切、PCR和序列测定结果显示,O/China/99株全基因组由8 200个核苷酸组成,构建的感染性cDNA与原毒株的核苷酸序列同源性为99.1%。结果表明,O/China/99缺失毒株全长cDNA分子克隆的构建成功。更多还原

【Abstract】 Two pairs of primers were designed and synthesized according to the genome sequence of deleted foot-and-mouth disease virus(FMDV) type O Pan-Asia strain O/China/99.Two amplified fragments from the genome of O/China/99 strain and a artificially-synthesized 5′UTR sequence deleted PKs were cloned into pOK12 vector.The recombinant was identified by restriction enzyme digestion,PCR and sequencing,respectively.The whole genome of FMDV O/China/99 strains was 8 200 nt in length.The nuc-leotide sequence of the full-length cDNA shared 99.1% identities with the strain O/China/99.The results showed that the full-length cDNA clone of FMDV O/China/99 strain was constructed successfully.更多还原