中文摘要：

目的构建两种人NDRG2亚型原核重组体pPROEX—HTb—NDRG2并进行表达、纯化、浓缩和鉴定。方法根据人NDRG2基因序列设计合成特异引物，聚合酶链反应（PCR）扩增，亚克隆入6×His融合表达载体pPROEX—HTb，在大肠杆菌BL21中经异丙基-β—D-硫代半乳糖苷（IPTG）诱导融合蛋白的表达，并用Protino Ni—TED Resin进行纯化。结果构建的重组质粒经酶切后分别可见约1100bp大小的片段，与预期结果一致，基因测序结果表明序列正确。在IPTG的诱导下重组体分别表达出分子量约45kD的表达产物，Western blot结果证实重组6×His融合蛋白可与抗-NDRG2蛋白抗体发生特异性反应；并通过Protino Ni—TED Resin成功获得了纯化蛋白。结论两种人NDRG2亚型基因分别被亚克隆至6×His融合表达载体pPROEX—HTb中，并获得成功表达和纯化，为该蛋白的生物活性及抑瘤作用的研究奠定了基础。

英文摘要：

Objective To construct the prokaryotic recombinant vectors for two subgroups of human NDRG2 gene and to identify their expression and purification. Methods The coding sequence of two subgroups of human NDRG2 were amplified with specific primers and cloned into the 6 Histidine （6 His） fusion expression vectors pPROEX-HTb. Then the recombinant plasmids were transformed into E. coli BL21 and the expressions of 6 His fusion protein were induced by adding isopropylthiogalactoside （IFFG）. Then fusion proteins were purified by Protino Ni-TED Resin. Results Restriction digestion of the constructed recombinant plasmid revealed the existence of gene segments about 1100 bp in length which were in accordance with what had been expected and the results of sequencing showed that the coding sequence of two subgroups of human NDRG2 gene were correct. Two protein bands with the molecular weight of about 45 kD were induced by IPTG in the recombinant plasmids. Western blot assay revealed that the 6 His fusion protein could be specifically recognized by NDRG2 antibody. And by Protino Ni-TED Resin, the fusion proteins were purified successfully. Conclusion The coding sequence of two subgroups of human NDRG2 gene have been successfully cloned into the 6 His fusion expression vectors pPROEX-HTb and efficiently expressed and purified, which lay the foundation for study on biological activity and the suppressing effect on tumor.