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目的 构建人白细胞介素 - 15 (IL- 15 ) c DNA真核表达载体 ,以期在哺乳类细胞中获得高效、稳定表达。方法 从外周血粘附性单核细胞 (PBMC)中提取总 RNA,经 RT- PCR方法获取了人 IL- 15 c DNA,并与 pc DNA3.1质粒的 Eco RI和 XbaΙ位点定向连接 ,构建受控于人巨细胞病毒 (CMV)启动子的重组真核表达载体 pc DNA3.1- IL - 15。结果 经 PCR扩增鉴定和 DNA序列分析 ,证明 IL- 15已经正确插入克隆载体且序列正确。
Objective To construct eukaryotic expression vector of human interleukin - 15 (IL - 15) c DNA for efficient and stable expression in mammalian cells. Methods Total RNA was extracted from peripheral blood mononuclear cells (PBMCs). Human IL-15 c DNA was obtained by RT-PCR and ligated with Eco RI and Xba I sites of pcDNA3.1 plasmid to construct The recombinant eukaryotic expression vector pc DNA3.1-IL-15, which is controlled by human cytomegalovirus (CMV) promoter. The results of PCR amplification identification and DNA sequence analysis showed that IL-15 has been correctly inserted into the cloning vector and the correct sequence.