中文摘要：

目的：原核表达并分离纯化人细胞骨架调节蛋白基因Nelin。方法：利用DNAstar软件预测Nelin的B细胞表位，选择Nelin基因B细胞表位较多、特异性最强的片段。用DNA重组技术，将此片段插入原核表达载体pE128a（＋），构建C末端带有His6标签的原核表达载体pE128a-Nelin，转化大肠杆菌B121（DE3），IPTG诱导，产物用Ni^2＋金属螯合吸附层析柱纯化。结果：成功构建了pE128a-Nelin表达载体，IFTG诱导后表达分子量约为17000的可溶性蛋白，Nelin-His6蛋白的表达量占菌体总蛋白的18％，经Ni^2＋金属螫合层析柱纯化，得到了电泳纯的Nelin—His6蛋白。结论：建立了稳定的Nelin基因的表达体系，为其功能研究奠定了基础。

英文摘要：

Objective：To purify recombinant protein of human cytoskeleton regulation gene Nelin expressed in Escherichia coll. Methods：The B cell epitopes of human Nelin gene were predicted by DNAstar software. A cDNA fragment of Nelin gene with two epitopes was amplified by PCR, and expression plasmid vector pg128a（ ＋ ） was ligated together with T4DNA ligase after digested by corresponding restriction endonucleases respectively. The new constructed vector pET28a-Nelin was identified by endonuclease digesting and confirmed by DNA sequencing. The BL21 （DE3） containing pET28a-Nelin plasmid was induced by IPTG, and the expressed protein were purified with Ni^2＋ metal chelate affinity chromatography columns. Results ：The recombinant expression vector pET28a-Nelin was successfully constructed. After induction, a new protein band of approximately 17 000 appeared on SDS-PAGE as soluble protein, which accounted for 18% of the total bacterial protein. Purified protein was obtained by Ni^2＋ metal chelate affinity chromatography. Conclusion：The stability expression method of Nelin gene is set up, and the result lays a foundation for the study of Nelin gene function.