羊痘病毒P32蛋白基因的克隆及其杆状病毒表达载体的构建  

Cloning of the gene of capripoxvirus major antigen P32 and construction of its baculovirus expression vector

作　　者：郭巍[1] 陈杰[2] 李一经[1] 李晓成[2] 黄保续[2] 张燕霞[2] 吴发兴[2] 

机构地区：[1]东北农业大学预防兽医教研室,黑龙江哈尔滨150030 [2]农业部动物检疫所国家动物流行病学研究中心疫病监测室,山东青岛266032

出　　处：《中国预防兽医学报》2004年第4期270-272,共3页Chinese Journal of Preventive Veterinary Medicine

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 。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.

关 键 词：羊痘病毒 P32蛋白 克隆 Bac-to-Bac○R状病毒表达系统 

分 类 号：S858.25[农业科学—临床兽医学] Q78[农业科学—兽医学]