中文摘要：

目的利用T7噬菌体展示技术筛选与白介素-32（interleukin-32，IL-32）相互作用的蛋白。方法构建IL-32α的原核表达重组质粒，表达并纯化重组IL-32α蛋白（recombinantIL-32et，rlL-32α）；以纯化后的rIL-32a作为靶蛋白，应用，17噬菌体展示技术对人肝细胞eDNA文库进行筛选；对筛选到的阳性克隆进行DNA测序及计算机辅助分析；利用哺乳动物双杂交实验和免疫共沉淀实验对筛选到的阳性克隆进行验证。结果成功表达并亲和纯化了dL-32α蛋白；经过5轮筛选，投入和产出的噬菌体滴度比值达到稳定：同源性分析初步确定9个与IL-32α蛋白相互作用的蛋白；进一步实验证实IL-32α与这9个蛋白片段能发生相互作用。结论利用17噬菌体展示技术成功筛选到9个与IL-32α相互作用的蛋白，我们的发现为深入揭示IL-32的生物学功能奠定了研究基础。

英文摘要：

Interleukin-32（IL-32） is a recently-described proinflammatory cytokine. In this study, we attempt to identify proteins that interact with IL-32. Firstly, a recombinant plasmid of pET28a-IL-32αwas constructed and then introduced into E. coli BL21 （DE3） to express recombinant IL-32α （rlL-32α）. SDS-PAGE analysis showed that rlL-32a was mainly expressed in the periplasm of host bacteria as soluble form. Then the rlL-32α was purified by His-bind resin column. Using rlL-32α as target protein, we performed the T7 phage display assay for 5 rounds from human liver cDNA library. The phage plaques were detected by PCR, and the PCR products were then subjected to DNA sequencing. Computer-assisted analysis of DNA sequencing results revealed that 9 protein peptides might interact with rIL-32α. Furthermore, mammalian two-hybrid assays and co-immunoprecipitation assays confirmed the interaction between the 9 proteins peptides and IL-32α. Taken together, we successfully identify 9 proteins that interact with IL-32. Our finding provides a novel insight for fully understanding the role of IL-32.