【摘要】 应用PCR方法扩增羊痘病毒特异性蛋白P32蛋白的基因 ,将其克隆至pMD18_T载体 ,测定核酸序列后将P32基因克隆至Bac_to_Bac○R杆状病毒表达系统中转移载体pFastBac1和pFastBacHTa ,进而转化入含穿梭载体Bacmid的感受态细胞DH10Bac中 ,获得两种携带P32基因的重组转染质粒 :Bacmid_Bac1_P32 ,Bacmid_BacHT_P32 ,经PCR试验鉴定 ,获得的这两种重组转染质粒是携带P32基因的杆状病毒表达载体 ,可分别用于转染Sf₉昆虫细胞以获得单一重组P32蛋白和六聚组氨酸融合体重组P32蛋白更多还原

【Abstract】 The P32 gene of capripoxvirus was multiplied by PCR and cloned into the pMD18T vector.After sequencing the gene,the P32 gene was separately cloned into the Bac-to-Bac^○R Baculovirus Expression System transfer vector pFastBac1 and pFastBacHTa,then the recombinant vectors were transfered into E.coli DH10Bac,which contains baculovirus shuttle vector Bacmid.We got two recombinant bacmids:Bac-Bac1-P32 and Bac-BacHT-P32.The two recombinant bacmids were approved to contain P32 gene by PCR test,so they shall be used to transfect Sf-9 insect cells,and then we shall separately get single recombinant P32 protein and recombinant 6-polyhistidine-P32 protein.更多还原