Construction and Expression of a Eukaryotic Vector for miR-145

Objective To construct a eukaryotic expression vector for miR-145,so as to lay a foundation for exploring its biological function in colon cancer.Methods The gene fragment of miR-145 was obtained by PCR and inserted into a eukaryotic expression plasmid,pCMV-myc.The constructed plasmid,pCMV-miR-145,was transfected into the colon cancer cell line HCT116.The expression level of miR-145 was detected by real-time PCR.Results The successful cloning of miR-145 into the eukaryotic expression vector pCMV-myc was confirmed by enzyme digestion and DNA sequencing analysis.The reconstructed plasmid could significantly increase the expression of miR-145 in HCT116 cells.Conclusions A eukaryotic expression vector for miR-145,pCMV-miR-145,is successfully constructed.It can highly express miR-145.