鸡肿瘤坏死因子-α原核表达及生物活性鉴定  

Chicken TNF-α prokaryotic expression and biologic activity

作　　者：王黎霞[1,2] 王彩霞[1] 张建军[1] 阮文科[1] 徐赓[1] 安健[1] 

机构地区：[1]北京农学院动物科学技术学院,北京昌平102206 [2]北京农业职业技术学院畜牧兽医系,北京房山102442

出　　处：《中国兽医杂志》2011年第11期6-9,共4页Chinese Journal of Veterinary Medicine

基　　金：国家自然科学基金资助项目(31072128);北京市教委科技发展计划资助项目(KM20071002001)

摘　　要：为研究肿瘤坏死因子-α(TNF-α)免疫功能,从鸡盲肠扁桃体克隆TNF-α的CDs保守区全序列,与pGEX-6p-1载体连接构建原核表达质粒,转化大肠杆菌M109,IPTG诱导蛋白表达,经包涵体洗涤、溶解、复性、亲和柱层析获得纯化的融合蛋白,经Western-blot鉴定为鸡TNF-α融合蛋白;用CEF/VSV法和MTT法检测表明,该融合蛋白的抗病毒活性为1.2×26 U/mg,在5-6稀释度下能显著促进淋巴细胞增殖。The objective is to investigate the function of TNF-α.The complete sequence of TNF-α CDs was cloned by RT-PCR from chicken cecal tonsil,then linked to pGEX-6p-1 expression vector to form prokaryotic expression plasmid and transformed into BL21 competent cell.GST-ChTNF-α fusion protein was obtained by the ablution and solution,re-natured of the inclusion body and affinity chromatography of Sepharose 4B.The specific band was identified by Western blot analysis using GST antibody.The results indicated the protein was GST fusion protein.The activity of the fusion protein was detected by CEF/VSV cell line and MTT experiment.The result showed that the fusion protein could resist VSV and stimulate the proliferation of lymphocytes.

关 键 词：肿瘤坏死因子-Α 原核表达 生物活性 

分 类 号：Q291[生物学—细胞生物学]