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| 猪CD8β分子重组蛋白的表达、纯化与免疫活性鉴定 | |
| 引用本文： | 薛萍,孙晓林,王晓霞,陈国华,房永祥,景志忠.猪CD8β分子重组蛋白的表达、纯化与免疫活性鉴定[J].甘肃农业大学学报,2009,44(4). |
| 作者姓名： | 薛萍 孙晓林 王晓霞 陈国华 房永祥 景志忠 |
| 作者单位： | 1. 甘肃农业大学动物医学院,甘肃,兰州730070;中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,甘肃省动物寄生虫病重点实验室,甘肃,兰州730046 2. 甘肃农业大学动物医学院,甘肃,兰州730070 3. 中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,甘肃省动物寄生虫病重点实验室,甘肃,兰州730046;西北农林科技大学动物医学院,陕西,杨凌712100 4. 中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,甘肃省动物寄生虫病重点实验室,甘肃,兰州730046 |
| 基金项目： | 国家高新技术"863"项目，甘肃省支撑计划项目 |
| 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. |
| 关 键 词： | 猪CD8β 原核表达 纯化 活性分析 |
Expression,purification and characterization of immunological activity of porcine CD8β protein |
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| XUE Ping,SUN Xiao-lin,WANG Xiao-xia,CHEN Guo-hua,FANG Yong-xiang,JING Zhi-zhong.Expression,purification and characterization of immunological activity of porcine CD8β protein[J].Journal of Gansu Agricultural University,2009,44(4). | |
| Authors: | XUE Ping SUN Xiao-lin WANG Xiao-xia CHEN Guo-hua FANG Yong-xiang JING Zhi-zhong |
| Institution: | XUE Ping1,2,SUN Xiao-lin1,WANG Xiao-xia2,3,CHEN Guo-hua2,FANG Yong-xiang2,JING Zhi-zhong1,(1.College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China,2.Key Laboratory of Veterinary Parasitology of Gansu Province,Key Laboratory of Veterinary Public Health of Ministry of Agriculture,State Key Laboratory of Veterinary Etiological Biology,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,3.College of Veterinary Medicine,Northwest Agric... |
| Abstract: | 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 the specific antibody against CD8β molecular,the antibody titre was higher than 1∶6 400 on average. |
| Keywords: | porcine CD8β(pCD8β) prokaryotic expression purification active analysis |
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