摘要
目的：鉴定构建的针对人核糖核酸酶抑制因子（ RI）的shRNA逆转录病毒载体。方法用脂质体法将针对人RI的shRNA逆转录病毒载体转染到人肝癌细胞SMMC7721中，用800 mg/L G418培养3～4周筛选产生稳定的细胞克隆，用RT-PCR和Western blot检测细胞中RI mRNA和蛋白的表达。结果 RT-PCR 和Western blot 表明，RI表达明显下调（ P＜0．05）。结论成功构建了针对人RI的shRNA逆转录病毒载体。
Objective To identify the silencing effect of the retroviral vector expressing shRNA targeting human ribonuclease inhibitor （ hRI） gene in the SMMC7721 cells .Methods The vector was transfected into SMMC 7721 cells with CellfectinR .After 3-4 weeks of selection of 800 mg/L G418, the expression of RI mRNA and protein was detected by RT-PCR and Western blotting in the SMMC7721 cells.Results Both RT-PCR and Western blotting showed that the expres-sion of hRI was significantly reduced in the SMMC 7721 cells transfected with shRNA-hRI retroviral vector as compared with the blank control group （P〈0.05）.Conclusion The retroviral vector of shRNA targeting hRI is successfully construc-ted.
出处
《山东医药》
CAS
2014年第13期13-15,共3页
Shandong Medical Journal
基金
辽宁省教育厅科研课题(L2011219)
大连大学本科生创新课题(2013108)
