Objective:To construct the eukaryotic expression vector of pEGFP-CD2AP and detect whether it could express in COS-7 cells. Methods:The coding sequence of CD2AP was obtained by PCR,then the PCR product was insert into pGEM T-Easy vector. After DNA sequence analysis,CD2AP was subcloned into pEGFP-C2 vector. Finally,a eukaryotic expression vector was obtained. The pEGFP-CD2AP expression vector was transfected with lipofectamine 2000 to COS-7 cells,and the expression of pEGFP-CD2AP was determined by GFP fluorescence. Results:The pEGFP-CD2AP expression vector could express both EGFP reporter gene and CD2AP in COS-7 cells. Conclusion:The pEGFP-CD2AP expression vector was constructed successfully,and it could be expressed in COS-7 cells. This work provided foundations for further study on the function of CD2AP.