新城疫病毒F基因在新型杆状病毒表达系统中的表达及重组病毒的免疫原性

Expression of Fusion Protein Gene of Newcastle Disease Virus in New Type Expression System of Baculovirus and Immunogenicity of the Recombinant Baculovirus

将新城疫病毒 (四平株 ) F基因插入到 p Fast Bac 质粒中 ,构建了重组质粒 p FF。 p FF转化大肠杆菌 DH10 Bac后 ,F基因在助手质粒 (helper plasmid)提供转座酶的情况下 ,通过转座进入 Bacmid,构建了重组 Bacmid(re- Bacmid)。在含有 X- gal/ IPTG、庆大霉素、卡那霉素和四环素的琼脂平板上 ,筛选白色菌落后 ,提取 re- Bacm id转染 Sf- 9昆虫细胞。在昆虫细胞内 ,re- Bacm id经复制、表达、装配 ,形成重组杆状病毒 ,并表达 F蛋白。经 SDS- PAGE和 Western- blot分析 ,表达的 F蛋白的分子大小为 6 30 0 0 ,并证明在昆虫细胞内表达的蛋白能部分糖基化。表达的 F蛋白约占细胞总蛋白的 10 %。动物试验证明 。

A recombinant transposon vector(pFF) was constructed by inserting the F gene of strain Siping into the transposon vector pFastBacⅠ.Then,pFF was transformed into E.coli DH10Bac containing bacmid and helperplasmid.Transposition was carried out and recombinant bacmid was constructed in the E.coli.The recombinant bacmid was transfected into sf 9 cells to generate recombinant baculovirus expressing F protein of NDV.The results of SDS PAGE and Western blot showed that the molecular weight of the expressed F protein was 63×10 3.The chickens were immunized with the recombinant baculovirus and antibody against F protein was determined by ELISA.The results showed that the F protein expressed by recombinant baculovirus could induce higher ELISA antibody titer against F protein,and had a good immunogenicity.