Construction of eukaryotic expression vector of human DR5 and establishment of its stable transfected NS-1 cell line

Objective: To construct eukaryotic expression vector of human death receptor(DR5) and transfect NS-1 cells to establish stable NS-1 cell line. Methods: The full-length DR5 cDNA fragment was amplified by RT-PCR from the human Jurkat cells and was inserted into eukaryotic expression vector pcDNA3.0.After the identification by digestion and sequencing on the recombinant eukaryotic expression vector pcDNA3.0/DR5,the recombinant was transfected into NS-1 cell by lipofectamine 2000.After screening culture by G418,stable transfected NS-1 cell line was established,and the transcription and expression of DR5 were identified by FACS. Results: The eukaryotic expression vector pcDNA3.0/DR5 was constructed successfully.The stable transfected NS-1 cell line was established.The DR5 protein was expressed successfully. Conclusion: The construction of the eukaryotic expression vector pcDNA3.0/DR5 and the establishment of stable transfected NS-1 cell line provide solid foundation for further experimental studies.