Construction and Identification of Prokaryotic Expression Vector of Human TNF-related Apoptosis Inducing Ligand

Objective:To clone and construct the prokaryotic expression vector of the TNF-related apoptosis inducing ligand gene(amino acids 114～281).Methods:RNA was extracted from the peripheral blood of health adult.A pair of primers with restriction site EcoR I/Xho I was designed.The extracellular domain region of TRAIL gene was amplified by RT-PCR and then cloned into the prokary-otic expression vector pGEX-6P-1.The recombinants were confirmed by EcoR I/Xho I digestion,PCR,and DNA sequencing.Result:The extracellular domain region with a molecular size of 501 bp of TRAIL gene was amplified successfully.DNA sequencing showed that the recombinant plasmid was successfully constructed.Conclusion:A prokaryotic expression vector pGEX-6P-1/TRAIL was con-structed successfully,which provides theory for tumor apoptosis research.