中文摘要：

目的：分别构建杜氏盐藻葡萄糖-6-磷酸异构酶（DsGPI）绿色荧光蛋白表达载体和真核表达载体。方法：PCR扩增得到上、下游分别添加EcoRⅠ和BamHⅠ酶切位点的DsGPI基因,双酶切后与绿色荧光蛋白表达载体pEGFP-C1连接,得到重组载体pEGFP-C1-DsGPI,转染狗肾细胞MDCK,观察绿色荧光蛋白在细胞中的定位;同理将上、下游分别添加BamHⅠ和EcoRⅠ酶切位点的DsGPI基因与真核表达载体pcDNA3.1（＋）连接,得到pcDNA3.1-DsGPI,转染食管癌EC9706细胞,分别应用RT-PCR法和Westernblot法检测DsGPI mRNA和蛋白的表达。结果：DsGPI在MDCK细胞中定位于细胞核,在EC9706细胞中被有效表达。结论：成功构建了DsGPI的绿色荧光蛋白表达载体和真核表达载体。

英文摘要：

To construct Dunaliella salina glucose-6-phosphate isomerase （DsGPI） green fluorescent protein ex- pression vector and eukaryotic expression vector. Methods：The complete cDNA sequences of DsGPI was cloned into the plasmid pEGFP-C1 to create a recombinant plasmid pEGFP-C1-DsGPI by using BamH I and EcoR I sites and identified. Localization of the EGFP fusion proteins was examined by fluorescence microscopy after the recombinant plasmids were transfected in MDCK cells. Recombinant plasmids pcDNA3. 1 （ ＋ ）-DsGPI were constructed with the same method and transfected in EC9706 cells,DsGPI mRNA was detected by RT-PCR and DsGPI protein were examined by Western blot. Results ： EGFP fusion proteins were mainly localized in the nucleus area in MDCK cells and expression of DsGPI increased in EC9706 ceils. Conclusion： DsGPI green fluorescent protein expression vectors and eukaryotic expression vectors has been constructed successfully.