小鼠microRNA-150慢病毒过表达载体及抑制载体的构建及鉴定

目的 构建microRNA-150慢病毒过表达载体及抑制载体.方法 设计针对microRNA-150前体的过表达基因片段及针对microRNA-150成熟体的反义片段,应用基因重组技术将目的 基因片段克隆到pWPI慢病毒载体中,并进行DNA测序鉴定重组克隆.结果 菌落PCR筛选鉴定出构建正确的慢病毒载体,DNA测序证实插入的基因序列正确.结论 成功构建microRNA-150慢病毒过表达载体及抑制载体,为研究microRNA-150在肝再生中的作用及其机制提供了稳定的细胞转染载体.

Construction and identification of lentlviral vector for microRNA-150 overexpression and inhibiton

Objective To construct and identify a lentiviral vector for microRNA-150 overexpression and inhibiton. Meth- ods The gene sequences of microRNA-150 precursor and mature microRNA-150 were chosen, and the overexpression sequences from microRNA-150 precursor and the antisense sequences from mature microRNA-150 were designed and linked into linearized pWPI vector. The recombinants were screened and identified by colony PCR and DNA sequencing. Results The target gene se- quences were successfully inserted into pWPI vector by identification of DNA sequencing. Conclusion The lentiviral microRNA overexpression vector and inhibitor vector has been successfully constructed, which provides a stable tool for further study of the role of mieroRNA-150 on liver regeneration.