【摘要】 目的 构建大鼠Wnt5a基因慢病毒并感染RSC96雪旺细胞,观察慢病毒感染情况及Wnt5a表达情况。方法 设计大鼠Wnt5a基因特异性RNAi序列,构建并包装成可感染细胞的慢病毒,感染RSC96雪旺细胞后通过Celigo观察细胞中荧光表达,并通过实时荧光定量聚合酶链反应(RT-qPCR)检测细胞中Wnt5a m RNA的表达情况。结果 构建的Wnt5a基因慢病毒在感染RSC96雪旺细胞后可表达绿色荧光,细胞中Wnt5a mRNA的平均表达量为0.206±0.056。结论 Wnt5a基因慢病毒构建成功,可为研究Wnt5a调控的周围神经损伤修复时雪旺细胞功能的相关研究提供便利。更多还原

【Abstract】 Objective To construct rat Wnt5a lentivirus and infect RSC96 Schwann cells,and to observe the infection and the expression of Wn5 a lentivirus.Methods The specific RNAi sequence of rat Wnt5a gene was designed,lentivirus that could infect cells were constructed and packaged.After RSC96 Schwann cells were infected,the fluorescent expression in cells was observed by Celigo,and the expression of Wnt5a mRNA in cells was detected by Real-time fluorescence quantitative polymerase chain reaction(RT-q PCR).Results The constructed Wnt5a lentivirus could express green fluorescence after infecting RSC96 Schwann cells.The average expression of Wnt5a mRNA in RSC96 Schwann cells was 0.206±0.056.Conclusion Wnt5a lentivirus was successfully constructed,to facilitate the studying on the function of Schwann cells in the repairing of peripheral nerve injury regulated by Wnt5a.更多还原