Objective To construct recombinant plasmid pCDH-Myc-UBR5 and then explore the biological function of ubiquitin protein ligase E3 component N-recognin 5 (UBR5) in regulating angiogenesis. Methods Using the complementary DNA (cDNA) of Michigan cancer foundation-7(MCF7) cells as a template, we divided the UBR5 gene coding region into two segments and amplified them by polymerase chain reaction (PCR). Then the two amplified fragments were inserted into the eukaryotic expression vector pCDH-Myc. Bacterial liquid PCR and DNA sequencing were adopted to identify the inserted fragments. The recombinant plasmid was tran-siently transfected into human embryonic kidney cells 293 SV40 LT(HEK293T) and the expression of Myc-UBR5 protein was detected by Western blotting. Co-immunoprecipitation (Co-IP) assay was employed to verify whether UBR5 interacts with dyskeratosis congenita 1 (DKC1). Results The recombinant plasmid pCDH-Myc-UBR5 was successfully constructed. The amplified sequence of UBR5 was successfully inserted into pCDH-Myc vector and expressed in HEK293T cells, which was confirmed by bacterial liquid PCR and DNA sequencing. The results of Co-IP assay indicated the interaction between UBR5 and DKC1. Conclusion The plasmid pCDH-Myc-UBR5 is successfully constructed by molecular cloning and expressed correctly in cells. UBR5 interacts with the angiogenic regulatory protein DKC1.