【摘要】 目的:研制hIL-17F/His重组蛋白并初步研究其体外生物学活性。方法:RT-PCR法克隆得到hIL-17F基因序列。将测序正确的hIL-17F基因片段装入PQE3.0原核表达载体构建重组载体hIL-17F/PQE3.0。将该重组载体导入宿主菌M15,经异丙基B-D硫代半乳糖苷(IPTG)诱导后产生hIL-17F/His重组蛋白,并用Western blot实验确认。结果:原核表达的hIL-17F/His重组蛋白变性、复性后,经过HiTrapTM亲和层析得到纯品蛋白。体外活性实验表明,该融合蛋白具有上调人巨噬细胞TNF-α、IL-6等分子表达的作用,并促进He-La细胞的增殖。结论:制备了具有生物学活性的hIL-17F/His重组蛋白,为进一步研究该分子独特的生物学功能奠定了基础。更多还原

【Abstract】 AIM: To Prepare recombinant human IL-17F/His protein and investigate its biological activity in vitro.METHODS: The gene region of human IL-17F was cloned by RT-PCR.After identification by sequencing,the hIL-17F gene encoding function domain was cloned into expression plasmid PQE3.0 and transfected into E.coli M15.By the induction of Isopropyl-β-D-Thiogalacto-Pyranoside(IPTG),recombinant IL-17F/His protein was effectively expressed in E.coli M15.The recombinant protein was identified by Western blot.RESULTS: After renaturation and purification by HiTrapTM affinity column,the recombinant protein can up-regulate macrophages to secret TNF-α,IL-6 and other relative cytokines.It also promoted proliferation of HeLa cells in vitro.CONCLUSION: hIL-17F/His recombinant protein is of high biological activity,which can be used to make further study of its special characteristic.更多还原