【摘要】 根据定点突变原理获得了包含口蹄疫病毒P1、2A、3C、3D及部分 2B编码区的目的基因片段 ,经SpeⅠ和HindⅢ双酶切后 ,与经相同方法处理的杆状病毒转移载体 pFastBacHT连接 ,得到了重组质粒 pFB P12X3C3D。经酶切和测序鉴定后 ,将其转化入含穿梭载体Bacmid的感受态细胞DH10Bac ,经抗性及蓝白斑筛选 ,得到了含P12X3C3D多基因的重组穿梭载体 ,将其命名为Bacmid P12X3C3D ;提取其DNA并转染Sf9昆虫细胞 ,最后获得含口蹄疫病毒P12X3C3D多基因的重组杆状病毒。更多还原

【Abstract】 The multi-gene, P12X3C3D fragment, which contains the complete genes P1, 2A, 3C, 3D and part of 2B of foot-and-mouth disease virus, was amplified by the site mutation PCR. After being digested with SpeⅠand HindⅢ respectively, the gene was linked to the baculovirus transfer vector pFastBacHT. The results showed that the recombinant vector, pFB-P12X3C3D, was constructed successfully. Then the recombinant vector was transformed into DH10Bac E.coli, and the positive recombinant bacmid Bacmid-P12X3C3D was screened according to the resistant of Gmr and Kar. After the Sf9 insect cells were transfected with the DNA extracted from Bacmid-P12X3C3D, the recombinant baculovirus particles were obtained. The achieved baculovirus particles were identified by PCR. The results showed that the multi-gene, P12X3C3D, have been introduced into the recombinant baculovirus correctly.更多还原