【摘要】 通过PCR和RT-PCR扩增出目的基因,将FMDV P1和3C基因通过IRES串联,构建出重组腺病毒穿梭质粒,在BJ5173细菌中同源重组获得同源重组腺病毒质粒,转化XL1-Gold超级感受态细胞以扩大培养,PacⅠ酶切后转染AD-293细胞,纯化获得了重组腺病毒。该重组腺病毒于AD-293细胞连续传代培养,初步浓缩后TCID50为1010.71/mL。应用O型口蹄疫病毒阳性血清进行间接荧光抗体试验,病毒感染的AD-293细胞可见清晰荧光,证明该重组腺病毒对目的基因进行了成功的表达,为FMDVP1/3C基因共表达腺病毒活载体疫苗的研究奠定了基础。更多还原

【Abstract】 The recombinant shuttle plasmid pShuttle-P1-IRES-3C was constructed successfully after the FMDV P1 and 3C genes were linked by IRES gene.Then the recombinant shuttle plasmid was linearized by PmeⅠand then transformed to BJ5183 cells containing the pAdEasy-1 bone plasmid,The homologous recombinant plasmid was transformed to XL-10 GOLD ultracompetent cells in order to obtain this plasmid in a large scale.A sufficient amount of purified recombinant adenovirus plasmid was digest with PacⅠ, and then transfected AD-293 cells using lipsome. After viral stock was passaged,the titer TCID50 was 1010.71/ml.Obvious fluorescence was observed in the AD-293 cell stained with standard serum of FMDV serotype O by IFA,which showed that the objective genes were expressed successfully.The results lay a foundation for the study of FMDV vaccine.更多还原