鸡Mx蛋白在293T细胞中的表达及其抗病毒活性

Research on the Expression Chicken Mx Protein in 293T Cell and Its Antiviral Activity

[目的]研究鸡Mx蛋白在293T细胞中的表达并检测其抗病毒活性。[方法]将鸡的Mx基因克隆入pEGFP-C1载体,经筛选鉴定后磷酸钙法转染293T细胞,并对转染细胞进行荧光分析以及RT-PCR与Western blot鉴定,同时提取细胞蛋白质,微量细胞病变试验检测其抗新城疫病毒的活性。[结果]试验构建的pEGFP-C1-cMx真核表达载体转染293T细胞后,在胞浆中可检测到绿色荧光,RT-PCR与Western blot结果进一步证实,pEGFP-C1-cMx可在293T细胞中表达,微量细胞病变试验显示,表达的融合蛋白可保护鸡胚成纤维细胞免受新城疫病毒感染。[结论]试验构建的pEGFP-C1-cMx表达载体可在293T细胞中表达具有抗新城疫病毒活性的鸡Mx蛋白。

[Objective] The expression of chicken Mx protein in 293T cell and the test of its antiviral activity were studied.[Method] The chicken Mx gene was cloned into pEGFP-C1 vector,and 293T cell was infected by it with calcium phosphate after the identification of the vector,and by screening of trans into,and the infected cell was analyzed in condition fluorescence and identified with RT-PCR and Western blot.Meanwhile the activity of its anti-Newcastle disease virus was tested through the micro-cytopathic test.[Results] The green fluorescence in the cytoplasm of 293T cell,which was infected with the eukaryotic expression vector constructed with pEGFP-C1-cMx,could be detected,and RT-PCR and Western blot results further confirmed that the pEGFP-C1-cMx could express in 293T cell and the micro-cytopathic test indicated that the fusion protein might protect cells into the chicken embryo fibroblasts from the infection of Newcastle disease virus.[Conclusion] The expression vector constructed with pEGFP-C1-cMx could express in 293T cell,which had the chicken Mx protein with the activity of anti-Newcastle disease virus.