Construction and expression of recombinant fusion of transmembrane programmed death ligand 1 and DsRed2

AIM: To construct the fusion gene of transmembrane programmed death ligand(PD-L1)and red fluorescent protein(DsRed2), and to study the expression and effect of the fused PD-L1/pDsRed2-N1. METHODS: The cloning technology was employed to construct the PD-L1/pDsRed2-N1 fusion gene. The fusion gene was transfected into NIT cells by lipofectamine. The expression of the fusion gene was determined by flow cytometry(FCM)and inverted phase contrast microscope. The effects on allo-spleen cells proliferation were detected by single mixed lymphocyte reaction. FCM was used to detect the proliferation of spleen cells. RESULTS: The expression of the fusion gene was observed in transiently transfected NIT cells. The fusion protein was expressed on cell membrane. These data suggested that the red fluorescent protein tag did not interfere with the natural assembly and the biological activity of PD-L1 molecule. CONCLUSION: The PD-L1-RFP fusion gene is constructed successfully and the fusion protein has biological activity.