〔Abstract〕 Objective To construct eukaryotic expression vector and to explore the subcellular localization of GPR107 in eukaryotic cells, human G protein coupled receptor 107(GPR107) gene is cloned. Methods Firstly, the cDNA of human GPR107 gene was cloned by molecular cloning technique, and the recombinant vector pCMV-Tag2 B-GPR107 was constructed by eukaryotic expression vector pCMV-Tag2 B. The expression of GPR107 was detected by Western blot, and the localization of GPR107 in 293 T cells was observed by immunofluorescence labeling and laser confocal microscope. Results The cDNA of human GPR107 was confirmed by gene sequencing and the eukaryotic expression vector pCMV-Tag2 B-GPR107 was constructed successfully. The results of Western blot showed that compared with the non-transfection group, the expression of GPR107 in the transfection group was significantly up-regulated. Laser confocal microscopy showed that GPR107 was mainly expressed in the cytoplasm. Conclusion GPR107 is overexpressed in 293 T cells and localized in the cytoplasm.