中文摘要：

目的 在原核表达系统中表达脂多糖应激新分子编码的蛋白质.方法 PCR扩增Hlrg Cdna编码区,克隆入表达载体pcTAT中,构建成融合6His的表达载体pcTAT-lrg,转化E.coli BL21（DE3）,以IPTG诱导.表达产物用SDS-PAGE及凝胶光密度扫描分析,用Ni-NTA亲和层析柱纯化.结果 经过IPTG诱导4 h,表达出相对分子质量（Mr）约25 000的蛋白,占菌体总蛋白的51%.表达产物为可溶性的,用Ni-NTA亲和层析柱纯化的表达蛋白达到电泳纯.加入培养液中的纯化蛋白,可在30 min内进入HEK293细胞.结论 在E.coli中成功地高表达人Lrg融合蛋白,并对其进行初步纯化,为人Lrg功能的研究打下基础.

英文摘要：

Objective To expression the recombinant human lipopolysaccharide respond gene （Lrg） protein in E. coli. Methods The encoding region of human Lrg cDNA was amplified by PCR, then subcloned into the prokaryotic expression vector pcTAT to construct pcTAT-hLrg. 6His-TAT was fused with coding sequence of hLrg cDNA in-frame with E. coli BI21 （ DE3）. Results 6His- TAT-hLrg had been expressed under IPTG induction, the expressed product accounted for 51% of the total protein, and was soluble. The 6His-TAT-hLrg purified through Ni-NTA affinity chromatography column reached electrophoretical pure. The purified protein could be found in the cytoplasm of HEK293 cells within 30 minutes. Conclusion Human Lrg fusion protein was expressed in E. coli successfully and high-efficiency, and Lrg was preliminary purified through Ni-NTA column, Which laid the foundation for further study of the function of human Lrg,