Objective: To construct pRetro-On/LTβR vector and obtain LTβR-expressing Jurkat cells,and explore the role of LTβR in T cells apoptosis. Methods: LTβR cDNA was obtained by PCR amplification and subcloned into pRetro-On plasmid. Then E.coli was transformed by the recombinant plasmid and positive clones under LB-Amp medium screen were used for recombinant plasmid extraction and purification. The recombinant plasmid pRetro-On/LTβR was identified by NotⅠ and BamHⅠ double enzyme digestion and DNA sequencing. Jurkat cells,a kind of T cell line,was transfected by the recombinant plasmid with Lipofectamine 2000. LTβR expression in Jurkat cells was induced by doxycycline(DOX) and identified by Western blot analysis and flow cytometry. The apoptosis of Jurkat cells induced by interaction of LTβR with the ligand,LIGHT,was detected by the method of annexin-V-PI double-stain flow cytometry. Results: Double enzyme digestion and DNA sequencing results confirmed that LTβR cDNA was inserted correctly into pRetro-On plasmid. The results of Western blot analysis and flow cytometry confirmed the LTβR expression in Jurkat cells. Apoptosis of transfected Jurkat cells significantly increased after LIGHT stimulation. Conclusion: The pRetro-On/LTβR was successfully constructed,and LTβR could be expressed in Jurkat cells using the recombinant plasmid. The apoptosis of LTβR-expressing Jurkat cells can be induced through LIGHT-LTβR pathway.