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目的 构建小鼠miR-29c过表达重组慢病毒载体,并对表达产物进行鉴定.方法 应用PCR法从小鼠MLE-12细胞中提取pre-miR-29c基因,测序后将其克隆到pLenti-CMV-EGFP慢病毒载体,通过PCR筛选及测序对阳性克隆进行鉴定.将miR-29c过表达慢病毒载体转染至293T细胞,进行慢病毒的包装和滴度测定.以RT-PCR检测慢病毒转染MLE-12细胞中miR-29c的表达量.结果 构建miR-29c慢病毒表达载体Lv-miR-29c经PCR和测序鉴定正确,并获得滴度为5.65× 108 IU/ml的病毒浓缩液,转染MLE-12细胞后miR-29c基因成功过表达.结论 成功构建miR-29c过表达慢病毒载体,为进一步研究miR-29c的功能奠定实验基础.“,”Objective To construct mouse miR-29c overexpression recombinant lentiviral vector and to identify the expressed product.Methods The pre-miR-29c gene was amplified from the mouse genome by PCR.The amplified product was cloned into pLenti-CMV-EGFP lentiviral vector.The positive clones were identified by PCR screening and sequencing.The miR-29c overexpression lentiviral vector was transfected into 293T cells for lentivirus packaging and titer determination.The lentivirus was infected with MLE-12 cells and the expression of miR-29c was detected by real-time PCR.Results PCR and DNA sequencing assays demonstrated that the miR-29c lentiviral expression vector Lv-miR-29c was successfully constructed and the titer of virus solution was 5.65×108 IU/ml.The miR-29c gene was overexpressed in MLE-12 cells.Conclusion The construction of miR-29c overexpression lentiviral vector is successful,which will lay the groundwork for the further study of the function of miR-29c.