鸡缩胆囊素-33串联体的构建、原核表达及免疫原性研究

根据鸡缩胆囊素（choleystokinin，cck）-33基因的碱基序列及大肠杆菌高频密码子，设计了大肠杆菌表达系统偏爱的cck基因序列．将cck-33基因克隆至pRSETA质粒上，利用载体上的同尾酶构建cck基因四串联体并在大肠杆菌EcoliBL21中成功表达．将所表达的融合蛋白进行纯化后，以纯化的蛋白为免疫原制备油佐剂疫苗主动免疫胡须肉鸡．结果表明，CCK蛋白主动免疫肉鸡后，抗血清水平显著升高，P／N值远远大于2．

Studies on the Construction, Expression and Immunogenicity of Chicken Choleystokinin-33 Concatemer

The choleystokinin-33(cck-33) gene was synthesized based on the chicken(cck-33) sequence and E.coli codon bias.The cck-33 gene was inserted into plasmids pRSET A and its concatemer was constructed based on the fact that isocaudarner enzyme existed.The recombinant CCK protein was expressed in E.coli BL 21 and purified successfully.The purified protein was used as an antigen to prepare oil-emulsion vaccines and then to immunize Huxu chicken.The result indicated that the anti-serum P/N of chicken immunized with purified protein was higher than 2.