摘要
对猪CD8β(pCD8β)基因重组蛋白的表达纯化条件以及免疫活性进行了研究.结果表明:pET-28a-pCD8β重组菌诱导表达产物的分子量约为24 ku,与预计的分子量大小相符.包涵体形式表达的重组蛋白的纯化产物经免疫印迹分析和ELISA检测证实有良好的特异性反应;SDS-PAGE后的目的胶带以直接研磨法和电洗脱纯化法加佐剂分别免疫BALB/c小鼠后,ELISA检测抗原免疫效价均大于1∶6 400,说明本试验对pET-28a-pCD8β纯化的重组蛋白具有良好的免疫原性和反应原性,可以制备CD8β的单克隆抗体.
The bioactivity and function of pCD8β were researched. The results showed that a constructed vector pET-28a -pCD8β in BL21 cells was induced by IPTG, the target protein was 24 Ku in size, which was consisted with expected one. The fusion protein was purified by recovering from SDS-PAGE. Western blot and ELISA analysis confirmed that the recombinant protein had excellent immunogenicity. Meanwhile, mice immunized with recombinant protein could produce antibody titre was higher than 1：6 400 on average. the specific antibody against CD8β molecular, the antibody titre was higher than 1：6 400 on average.
出处
《甘肃农业大学学报》
CAS
CSCD
北大核心
2009年第4期4-9,共6页
Journal of Gansu Agricultural University
基金
国家高新技术"863"项目(2006AA10A203)
甘肃省支撑计划项目(0804NKCA076)
关键词
猪CD8β
原核表达
纯化
活性分析
porcine CD8β（pCD8β）
prokaryotic expression
purification
active analysis
