中国畜牧兽医2011，Vol.38Issue(11)：102-105,4.
猪白细胞介素-6的原核表达及其生物学活性研究
Prokaryotic Expression and Bioactivity of the Porcine Interleukin-6
摘要
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 urea, refolding by a self-innovative renaturation buffer and dialyzing by PBS buffer etc. The porcine IL-6 ELISA assay was used to detect the specifically immunological activity of the rPoIL-6 protein, and the cell proliferation activity induced by rPoIL-6 protein was assessed by MTS assay. The result showed that the PoIL-6 mature peptide gene with a length of 555 bp was successfully cloned, expressed, and purified with the molecular mass of about 20 ku and more than 95% pure on SDS-PAGE, which indicated the correct PoIL-6 fusion protein had been obtained. The rPoIL-6 protein could specifically react with McAb aginast rPoIL-6 and significantly promote the proliferation of porcine spleen lymphoblast cells.关键词
猪白介素-6/原核表达/纯化/活性分析Key words
porcine interleukin-6/ prokaryotic expression/ purification/ activity assay分类
生物科学引用本文复制引用
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):102-105,4.基金项目
河南省科技攻关计划项目资助(08210210008). （08210210008）
