摘要
根据牛瑟氏泰勒虫主要表面蛋白基因p23和p33的已知序列,设计特异性引物,将克隆产物与pVAX1表达载体连接,得到真核重组表达质粒pVAX1-p33-p23,脂质体介导其转染Hela细胞后进行表达产物的鉴定,最后进行BALB/c小鼠免疫试验。结果,目的基因在真核细胞内得到正确表达,动物免疫试验pVAX1-p33-p23核酸疫苗能够提高小鼠的细胞免疫和体液免疫水平,并且pVAX1-p33-p23免疫组的免疫效果均高于pVAX1-P33和pVAX1-P23免疫组(P<0.05)。从而证明,牛瑟氏泰勒虫的重组pVAX1-p33-p23核酸疫苗成功构建。
To construct the fusion expression vector of p33-p23 major surface protein genes from Theileria. sergenti,two pairs of primer were designed and synthetized according to P33 and P23 gene sequence of Theileria Sergenti,the cloning products were inserted into expression vector pVAX1. Hela cells were transfected with the obtained recombinant eukaryon reorganization vector pVAX1-p33-p23. The expressed product was identified by Western blot. Finally the BALB/c mice were used for immunising experiment. Result showed the goal gene was correctly expressed in the eukaryotic cell. The animal immunising experiment result indicated that pVAX1-p33-p23 can enhance mouse＇s cellular immunity and the humoral immunity level, and the immunity group＇s immunity effect of pVAX1-p33-p23 is higher （P〈0.05） than that of pVAX1-P33 and the pVAX1-P23 immunity group. Conclusion： Recombinant plasmid pVAX1-p33-p23 was successfully constructed.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2008年第10期1171-1173,1180,共4页
Chinese Journal of Veterinary Science
基金
吉林省科技发展计划项目(20050703-4)
