| 猪白细胞介素-6的原核表达及其生物学活性研究 |
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| 引用本文： | 马文涛,闫若潜,吴志明,刘光辉,盛敏,谢彩华.猪白细胞介素-6的原核表达及其生物学活性研究[J].中国畜牧兽医,2011,38(11):102-105. |
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| 作者姓名： | 马文涛 闫若潜 吴志明 刘光辉 盛敏 谢彩华 |
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| 作者单位： | 1. 河南农业大学牧医工程学院,河南郑州,450002
2. 河南省动物疫病预防控制中心,河南郑州,450008 |
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| 基金项目： | 河南省科技攻关计划项目资助(08210210008) |
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| 摘 要： | 本研究采用RT-PCR方法自细菌脂多糖(LPS)刺激的猪脾脏细胞总RNA中扩增、克隆猪白介素-6(porcine interleukin-6,PoIL-6)成熟肽基因,并亚克隆入pQE30载体进行原核表达,对表达的重组融合猪白介素-6(rPoIL-6)蛋白通过尿素变性、低浓度复性液复性、PBS透析等步骤进行复性、纯化,采用PoIL-6 ELISA试剂盒检测rPoIL-6蛋白与抗PoIL-6单抗发生特异性免疫反应的活性;采用MTS法检测rPoIL-6蛋白促猪脾脏细胞的增殖活性。结果表明,成功克隆了全长555 bp的PoIL-6成熟肽基因;表达的rPoIL-6蛋白分子质量大小约20 ku;纯化后的rPoIL-6蛋白纯度在95%以上,可和抗PoIL-6单抗发生特异性免疫反应,并且具有显著促猪脾脏细胞增殖活性。
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| 关 键 词： | 猪白介素-6 原核表达 纯化 活性分析 |
| 收稿时间： | 2011-03-22 |
Prokaryotic Expression and Bioactivity of the Porcine Interleukin-6 |
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| MA Wen-tao,YAN Ruo-qian,WU Zhi-ming,LIU Guang-hui,SHENG Min,XIE Cai-hua.Prokaryotic Expression and Bioactivity of the Porcine Interleukin-6[J].China Animal Husbandry & Veterinary Medicine,2011,38(11):102-105. |
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| Authors: | MA Wen-tao YAN Ruo-qian WU Zhi-ming LIU Guang-hui SHENG Min XIE Cai-hua |
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| Institution: | 1. College of Animal Husbandry and Veterinary Medicine,Henan Agricultural University,Zhengzhou 450002,China;2. Henan Center for Animal Diseases Control & Prevention,Zhengzhou 450008,China |
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| Abstract: | The porcine interleukin-6(PoIL-6) mature peptide gene was amplified from the total cellular RNA of porcine spleen cells induced by bacterial lipopolysaccharides(LPS) by RT-PCR,and the PCR production was subsequently cloned,sequenced and sub-cloned into the prokaryotic expression vector pQE30.The PoIL-6 mature peptide gene was expressed in E.coli JM109 and the expressed fusion recombinant protein(rPoIL-6) was purified under the innovated recombinant fusion protein purification method of denaturing by 8 mol/L... |
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| Keywords: | porcine interleukin-6 prokaryotic expression purification activity assay |
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