中文摘要：

通过构建人白细胞介素7的重组真核表达载体,转染HEK293细胞,实现IL-7在HEK293细胞中的稳定分泌表达,为生产试剂级的IL-7奠定基础。从质粒p LV-CMV-EF1-GP-r IL-7中扩增人IL-7全长基因,并将其构建于pc DNA3.1真核表达载体上。利用脂质体法将重组质粒转染HEK293细胞,经G418筛选及单克隆,获得稳定分泌表达IL-7的HEK293-r IL-7细胞系。成功构建真核表达质粒pc DNA3.1-r IL-7,并实现IL-7在HEK293细胞上的稳定分泌表达,最终获得了1株表达量较高的细胞株,命名为HEK293-IL7-2G3细胞,培养48 h的细胞上清中IL-7的含量为3.2 ng/m L。为进一步实现人白细胞介素7在哺乳动物细胞中的分泌表达及药物开发奠定基础。

英文摘要：

The objective of this study was to construct eukaryotic expression vector of human interleukin-7,and make it expressed in HEK293 cells. IL-7 gene was amplified by PCR from the lentivirus expression vector p LV-CMV-EF1-GPL-r IL-7,and then cloned into the pc DNA3. 1 vector to construct eukaryotic expression vector pc DNA3. 1-r IL-7. The constructed plasmid was transfected into the HEK293 cells via LipofectamineTM2000 separately and screened by G418 continuously. By G418 screening and monoclonal,stably secreting expression IL-7 cell line HEK293-r IL-7 was obtained,successfully constructed eukaryotic expression vector pc DNA3. 1-r IL-7,and expressed in HEK293 cells. By cell cloning,one cell line with high secretory expression was obtained,named HEK293-r IL7-2G3.The content of IL-7 in the supernatant after 48 h was 3. 2 ng/m L.