中文摘要：

目的：构建双表达人Arid5a（hArid5a）及报告基因eGFP的重组腺病毒载体，为研究Arid5a的生物功能打基础。方法：通过常规分子克隆方法及基因重组技术获得双表达hArid5a及eGFP的腺病毒载体。通过荧光显微镜观察eGFP的表达来确定病毒包装情况，Western blot检测Arid5a蛋白表达。结果：将hArid5a克隆到腺病毒穿梭载体中，获得pAd5-CMV-hArid5a 载体， 然后将Pac I线性化的pAd5-CMV-hArid5a与Pac I线性化的携带报告基因eGFP的病毒骨架通过基因重组，最终获得携带有报告基因eGFP及人Arid5a的腺病毒载体Ad5 CMV-hArid5a/eGFP。将纯化获得的腺病毒载体Ad5 CMV-hArid5a/eGFP感染U87细胞后，通过荧光显微镜观察eGFP的表达；通过Western blot检测到感染细胞中hArid5a表达。结论：成功构建了携带报告基因eGFP的hArid5a的腺病毒表达载体，为进一步研究hArid5a的生物功能奠定了基础。

英文摘要：

Objective： To construct and identify recombinant adenovirus dually expressing human AridSa （hAridSa） and reporter gene eGFP. Methods： The recombinant adenovirus dually expressing human AridSa （hAridSa） and reporter gene eGFP was constructed by conventional molecular cloning techniques. The cytopathic effect of recombinant adenovirus was evaluated by fluorescence microscope,and the expression of hAridSa was determined by Western blot. Results：hAridSa gene was amplified by PCR from cDNA library,and then cloned into adenovirus shuttle vector （ pad5 - CMV - hAridSa）. The adenovirus shuttle vector （ pAd5 - CMV - hAridSa） and adenovirus backbone carrying eGFP in the E3 were linearized by Pac I and co - transfected into HEK293 cells. Through further amplification and purification, recombinant adenovirus （AdS CMV -hAridSa/eGFP） expressing eGFP and hAridSa was successfully constructed. Green fluorescent protein expression from recombinant adenovirus was observed in infected cells by fluorescence microscope. The expression of hAridSa can be detected in the ceils infected with AdS CMV -hArid5 a/eGFP by Western blot. Conclusion：These results indicated that we have successfully constructed recombinant adenovirus dually expressing eGFP and hAridSa,which will provide a useful tool for the further study on the function of AridSa.