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目的 :构建人白细胞介素 - 15 (hIL - 15 )真核表达载体并在肝癌细胞BEL - 74 0 2中表达 ,为进一步研究IL - 15的功能及临床应用奠定基础。方法 :采用RT -PCR方法自正常成人外周血单核细胞中扩增出hIL -15cDNA ,将其克隆至T -vector中 ,经测序确证后 ,将该基因定向插入真核表达载体pcDNA3.1hisB中 ,脂质体法转染BEL - 74 0 2细胞进行表达 ,鉴定表达产物的生物学活性。结果 :测序结果与已报道hIL - 15cDNA序列一致。脂质体转染后获得 6个BEL - 74 0 2细胞阳性克隆 ,IL - 15活性测定为 15 6～ 2 5 2u/ml。结论 :成功地构建了真核表达载体 pcDNA3.1hisB -IL - 15 ,IL - 15cDNA转导的人BEL - 74 0 2肝癌细胞可表达有生物活性的IL - 15。
AIM: To construct eukaryotic expression vector of human interleukin - 15 (hIL - 15) and express it in hepatocellular carcinoma cell line BEL - 74 0 2, which will lay the foundation for further study of the function and clinical application of IL - 15. Methods: hIL-15 cDNA was amplified from normal adult peripheral blood mononuclear cells by RT-PCR and cloned into T-vector. The DNA sequence was inserted into eukaryotic expression vector pcDNA3.1hisB , And then transfected into BEL - 7402 cells by lipofectamine for expression. The biological activity of the expressed product was identified. Results: The sequencing results were consistent with the reported hIL - 15 cDNA sequence. Six BEL - 7402 positive clones were obtained after lipofectamine transfection. The activity of IL - 15 was determined to be 15 6 ~ 25 2 u / ml. CONCLUSION: The eukaryotic expression vector pcDNA3.1hisB-IL - 15 was successfully constructed and human BEL - 74 0 2 hepatoma cells transduced with IL - 15c could express bioactive IL - 15.