Objective To construct expression vectors that enhanced yellow fluorescent protein (EYFP) fused with Apelin receptor (Apelin-R) and Renilla reniformis (Rluc) fused with Neurotensin type 1 receptor (NTSR1), to investigate the heterdimerization of human Apelin-R and NTSR1 and its role in signal transduction.Methods The human NTSR1 gene was amplified by PCR using the plasmid pcDNA3.1-hNTSR1 as template.The PCR product was digested, ligased with the plasmid pRluc and then be transformed into the competent cell Top10.The construct was identified by DNA sequencing.The recombinant plasmid was transiently transfected into human embryonic kidney 293 (HEK293) cells, and the expression of pRluc-hNTSRI-pcDNA3.1 was detected by eonfocal microscopy and Western blot.pEYFP-hApelin-R was also constructed by the same mean.Results Two fragments of 1257 bp and 1143 bp were amplified by PCR, and the DNA sequences were identical with the gene in GenBank (NM_002531, NM_005161), respectively.Western blot showed 91 kDa and 69 kDa band.Confocal microscopy showed that NTSR1 and Apelin-R were expressed on the plasma membrane.Conclusion The pRlue-hNTSRI-pcDNA3.1 and pEYFP-hApelin-R eukaryotic expression vectors were successfully constructed, and the expression vector could be used to investigate the heterdimerization of human Apelin-R and NTSR1 and its role in signal transduction, which will provide new target for drug development.