Construction of Eukaryotic Expressing Vector of Human PD-L1 and Establishment of Stable Transfectant HEK293 Cell Line

Objective To construct eukaryotic expressing vector of human PD-L1 gene and transfect HEK293 cells so as to establish stable HEK293 cell line. Methods PD-L1 gene was amplified by polymerase chain reaction from the human heart cDNA library and confirmed by DNAsequence analysis. The PD-L1 gene was then digested with the restriction endonucleases Xho1 and EcoR1 and inserted into eukaryotic expressing vector pIRES2-EGFP. pIRES2-EGFP/PD-L1 was transfected into HEK293 cells by lipofectamineTM2000. After screening culture by G418, stable transfected HEK293 cells line was established, and the expression of PD-L1 was identified by FCM, RT-PCR and Western blot. Results The eukaryotic expressing vector pIRES2-EGFP/PD-L1 was constructed, stable transfected HEK293 cell line was established, and PD-L1 gene was expressed successfully. Conclusion The construction of eukaryotic expressing vector pIRES2-EGFP/PD-L1 and the establishment of stable transfected HEK293 cell line have provided solid experiment foundation for further studies on the function of PD-L1.