中文摘要：

目的：构建人水甘油通道蛋白9（aquaglyceroporin9,AQP9）重组质粒,并检测其融合蛋白在真核细胞COS-7中的表达,为阐明AQP9在非酒精性脂肪肝病（nonalcoholic fatty liver disease,NAFLD）中的作用奠定基础。方法：通过RT-PCR方法从人肝脏组织中获得AQP9基因,插入pEGFP-N1质粒,构建重组质粒pEGFP-N1-AQP9,转染COS-7细胞,通过荧光显微镜检测其转染情况,RT-PCR和Western blot检测AQP9在真核细胞中的表达。结果：PCR及酶切鉴定显示插入人AQP9大小正确。测序鉴定显示,1个碱基发生突变,45位：A→G,为无义突变。通过RT-PCR和Western blot检测到AQP9表达,说明AQP9基因在COS-7细胞中表达并与绿色荧光蛋白融合,且具有免疫原性。结论：成功构建了pEGFP-N1-AQP9重组质粒,并在COS-7细胞中表达,为进一步研究AQP9在NAFLD发生发展中的作用奠定基础。

英文摘要：

Objective：To construct human aquaglyceroporin 9（AQP9）recombinant plasmid and to detect its expressions in COS-7 cells in order to elucidate the role of AQP9 in pathogenesis of nonalcoholic fatty liver disease（NAFLD）.Methods：AQP9 gene was obtained by RT-PCR from human hepatic tissue and was cloned into pEGFP-N1 plasmid.pEGFP-N1-AQP9 recombinant plasmid was constructed,identificated and transfected into COS-7 cells.Transfection was detected by fluorescence microscopy;expressions of AQP9 were detected by RT-PCR and Western blot.Results：PCR and enzyme digestion analysis showed that AQP9 size was right.Equencing analysis found a nonsense mutation,45 th：A→G.RT-PCR and Western blot demonstrated that AQP9 was expressed in COS7 cells,suggesting that pEGFP-N1-AQP9 was expressed in COS-7 cells and had immunogenicity.Conclusions：pEGFP-N1-AQP9 recombinant plasmid is constructed and expressed successfully in COS-7 cells,which would lay a foundation for further study of AQP9 in the pathogenesis of NAFLD.