中文摘要：

目的在真核细胞中表达hIL-23（p19）／mFc融合蛋白并初步研究IL-23生物学特性。方法应用RT-PCR法克隆获得hIL-23（p19）／mFc基因片段。将测序正确的hIL-23（p19）／mFc序列插入pCEP4质粒构建pCEP4／hIL-23（p19）／mFc真核表达载体。转染人肾上皮293T细胞后，筛选阳性表达细胞株。RT-PCR和Western blot法鉴定hIL-23（p19）／mFc基因和蛋白表达。结果成功构建了pCEP4／hIL-23（p19）／mFc重组表达载体，并在293T细胞中稳定表达。获得的hIL-23（p19）／mFc重组蛋白能促进人T细胞hIL-17和hIL-10的表达。体外刺激巨噬细胞，能显著增加TNF-a等炎症因子的基因表达。结论成功建立稳定表达hIL-23（p19）／mFc重组蛋白的293T细胞系，可用于进一步研究hIL-23的生物学功能。

英文摘要：

Objective To construct pCEP4/human interleukin-23 （ hIL-23 ） （ p19 ）/mFc recombinant expression vector and express it stably in eukaryotic cells. For studying the biological activity in vitro. Methods The CDS region of hIL-23 （p19）/mFc gene was cloned by RT-PCR. After identification by sequencing, the hIL-23 （plg）/mFe gene was inserted into the expression vector of pCEP4 to construct the recombinant vector pCEP4/hIL-23 （p19）/mFc, and then transfected into 293T cells. The transgenic 293T cell line stably expressing rhIL=23 （plg）/mFc protein was selected in the presence of hygromycin B. After FCS-free cultivation and sub-cloning, the IL-23 （p19）/mFc gene and protein expression were confirmed by RT-PCR, ELISA and Western blot analysis. Results The recombinant pCEP4/hIL-23（p19）/mFc and its transgenic 293T cells stably expressing rhIL-23（plg）/ mFc protein were obtained successfully. Flow cytometry analysis showed that it could stimulated human T cells to excrete hIL-10 and hIL-17 and RT-PCR analysis exhibited high TNF-α mRNA expression on human macrophages in vitro. Conclusion The recombinant pCEP4/hIL-23 （p19）/mFc and its transgenic 293T cells stably expressing rhIL-23 （p19）/mFc protein have been obtained successfully, which can be used to analyze the biological functions of hIL-23.