Abstract: Objective：To construct the pQCXIH-HA-FBXL11 retroviral expression plasmid, identify the expression, and establish stable cells that over-expressed FBXL11 in mesenchymal stem cells (MSCs). Methods：PCR was used to amplify the target gene, FBXL11, using cDNA of the dental pulp stem cells as a template. PCR products were purified and recycled, ligation with appropriate vector. Enzyme digestion and sequencing were used to identify the pQCXIH-HA-FBXL11 plasmid. The plasmids were transfect in 293T cells using FuGENE6, Western blot and real-time PCR were used to detect the expressions of FBXL11 at the protein and mRNA levels level. Retroviruses were infected the dental pulp-derived mesenchymal stem cells (DPSCs). Results：We successfully constructed the retroviral plasmid of pQCXIH-HA-FBXL11, confirmed its expression in 293T cells by Western blot and real-time PCR, and successfully established the FBXL11 over-expressed stable cells in DPSCs. Conclusions：The construction and expression of retroviral pQCXIH-HA-FBXL11 and over-expressed FBXL11 in DPSCs were the basis to investigate the function of FBXL11 in mesenchymal stem cells.