人重组IL-17/His蛋白的原核表达、纯化及其生物学活性

Prokaryotic expression,purification and biological activity of recombinant human IL-17/His protein

目的:研究人IL-17的体外生物学活性。方法:采用RT-PCR的方法克隆得到了hIL-17基因序列。将测序正确的人IL-17基因装入PQE3.0原核表达载体构建重组载体hIL-17/PQE3.0。该重组载体导入宿主菌M15,经异丙基β-D硫代半乳糖苷(IPTG)诱导产生IL-17/His融合蛋白,并经West-ernblot实验确认。结果:原核表达的hIL-17/His重组蛋白经变性、复性后,利用HiTrapTM亲和层析得到纯品蛋白。体外活性实验表明,该融合蛋白具有刺激人宫颈癌细胞株HeLa分泌IL-6和GM-CSF的作用。结论:制备了具有生物学活性的IL-17/His重组蛋白,为进一步研究该分子在自身免疫疾病等方面的作用奠定了基础。

AIM： To investigate the biological activity of recombinant human IL-17 protein in vitro. METHODS： The gene region of human IL-17 was cloned by RT-PCR. After identification by sequencing, the hlL-17 gene encoding function domain was cloned into expression plasmid PQE3.0 and transfect into E. coli M15. By the induction of Isopropyl- 13-D-Thiogalacto-Pyranoside （ IPTG）, recombinant IL-17/His protein was effectively expressed in E. coil M15. The recombinant protein was identified by Western blot. RESULTS： After denaturation, renaturation and purification by HiTrapTM affinity column, the recombinant protein stimulated HeLa, a human uterine cervix cancer cell line, to excrete IL-6 and GM-CSF in vitro. CONCLUSION： IL-17/His reconbinant protein is of high biological activity, which can be used to make further study of auto-immune diseases.