目的构建小鼠胰腺β细胞NIT-1中miR-125b慢病毒过表达载体。方法从小鼠基因组DNA中用PCR扩增miR-125b的前体序列,经NheⅠ和PstⅠ酶切后连接到Lv-CMV-GFP载体中,并对重组质粒进行双酶切鉴定和测序分析。将miR-125b过表达载体、pVSV-G和delta8.91质粒共转染293T细胞,收获慢病毒感染NIT-1细胞,流式细胞术分选阳性细胞,RT-PCR鉴定稳转细胞株中miR-125b的表达。结果成功构建了miR-125b慢病毒表达载体,感染小鼠胰腺β细胞NIT-1后能够成功过表达miR-125b。结论成功构建miR-125b慢病毒过表达载体,为进一步研究其功能建立了实验基础。 Objective To construct miR-125b lentivirus overexpression vector in mouse pancreatic β-cell NIT-1. Methods The precursor sequence of miR-125b was amplified by PCR from mouse genomic DNA and ligated into Lv-CMV-GFP vector after digestion with NheⅠ and PstⅠ. The recombinant plasmid was identified by double enzyme digestion and sequencing analysis. The miR-125b overexpression vector, pVSV-G and delta8.91 plasmids were co-transfected into 293T cells, lentivirus-infected NIT-1 cells were harvested, and the positive cells were sorted by flow cytometry. RT-PCR was used to identify the miRs -125b expression. Results The miR-125b lentiviral vector was successfully constructed and miR-125b was overexpressed successfully after infection of mouse pancreatic β-cell NIT-1. Conclusion The miR-125b lentivirus over-expression vector was successfully constructed and established the experimental basis for further study of its function.