鸡α-干扰素成熟肽的原核表达及其多价血清的制备  

Prokaryotic expression of chicken α-interferon mature peptide and its preparation of polyvalent serum

作　　者：王永娟[1] 朱善元[1] 左伟勇[1] 韩小英[1] 吴双[1] 王安平[1] 洪伟鸣[1] 黄文强[1] 

机构地区：[1]江苏畜牧兽医职业技术学院/江苏省兽用生物制药高技术研究重点实验室,江苏泰州225300

出　　处：《扬州大学学报（农业与生命科学版）》2013年第1期32-35,40,共5页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：江苏省教育厅科技产业化推进项目(JHZD09-71);江苏畜牧兽医职业技术学院博士启动基金项目(ZD200908);江苏畜牧兽医职业技术学院重点支持项目(ZD1204)

摘　　要：以PCR方法从活化的鸡外周血淋巴细胞中扩增鸡α-干扰素的成熟肽基因序列,构建原核表达质粒pET32a-α和pGEX6p-α;SDS-PAGE及Western blotting法检测两载体特异性表达成熟肽;将纯化的pET32a-α诱导表达产物免疫小鼠,收获小鼠血清并以pGEX6p-α诱导表达产物检测其中特异性抗体滴度。结果表明:扩增的鸡α-干扰素成熟肽基因成功表达,原核表达产物免疫小鼠可产生特异性抗体,抗体效价为1∶12 000。其结果为制备安全、高效的重组干扰素奠定了基础,为定量检测机体内产生的α干扰素进而了解机体免疫状态提供较为可靠的技术手段。Mature peptide of chicken IFN-a gene was amplified from activated chicken peripheral blood lymphocytes by PCR. pET32a-α and pGEX6p-α were constructed for IFN-α expression. Purified pET32a-α induced protein was injected into mice for peptide-specific antibody which was detected by pGEX6p-α induced protein. As a result, amplification of mature peptide of chicken IFN-α gene could be successfully expressed in prokaryotic expression vector for immunized mice to produce specific antibody, the antibody titer was 1 ： 12 000. The results provide the foundation for the preparation of safe and efficient recombinant interferon and a more reliable technique for the quantitative detection of the body producing IFN-α which can reflect the immune status of the body against pathogens, as well as a detection method for some other biomolecules with no commercial antibodies.

关 键 词：鸡 Α-干扰素 原核表达 抗体 

分 类 号：S858.31[农业科学—临床兽医学]