[Construction and expression of recombinant fusion of transmembrane programmed death ligand 1 and DsRed2].
OBJECTIVE
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.


CONCLUSIONS
The PD-L1-RFP fusion gene is constructed successfully and the fusion protein has biological activity.
Liao
Wen-jun
WJ
No.455 Hospital of PLA, Nephrology Center of Nanjing Military Area, Shanghai 200052, China. wenjun_liao@hotmail.com
Sun
Jing
J
Zhu
Hui-fen
HF
Zhang
Jin-yuan
JY
Shen
Guan-xin
GX
D000818:Animals; D000954:Antigens, Surface / Q000235:genetics / Q000378:metabolism*; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D050417:Insulin-Secreting Cells / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008858:Microscopy, Phase-Contrast; D011247:Pregnancy; D061026:Programmed Cell Death 1 Receptor; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D000097573:Red Fluorescent Protein
D000954:Antigens, Surface; D051017:Apoptosis Regulatory Proteins; D008164:Luminescent Proteins; D061026:Programmed Cell Death 1 Receptor; D011993:Recombinant Fusion Proteins; D000097573:Red Fluorescent Protein; C491383:Pdcd1 protein, mouse