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目的:获得TRAIL全长基因并在原核表达系统中表达。方法:通过RT-PCR方法,从人外周血细胞中克隆TRAIL全长基因,再克隆其至原核表达载体pGEX-2T中,并在大肠杆菌JM109细胞中表达。结果:获得了TRAIL基因。序列分析表明,该序列与GenBank数据库中的序列一致。其原核表达产物能够抑制肿瘤细胞系KB细胞及Hela细胞的生长,最终导致细胞凋亡。结论:原核表达体系表达了具有生物学活性的TRAIL,为进一步研究其抗肿瘤活性奠定了基础。
Objective: To obtain full-length TRAIL gene and express it in prokaryotic expression system. Methods: The full-length TRAIL gene was cloned from human peripheral blood by RT-PCR and cloned into prokaryotic expression vector pGEX-2T and expressed in E. coli JM109. Results: The TRAIL gene was obtained. Sequence analysis showed that the sequence was consistent with the sequence in GenBank database. Its prokaryotic expression product can inhibit the growth of tumor cell lines KB cells and Hela cells, eventually leading to apoptosis. CONCLUSION: The prokaryotic expression system expresses the biologically active TRAIL, which lays the foundation for further study of its anti-tumor activity.