人IL-6受体胞外区真核表达载体的构建及体外表达

目的构建人IL-6受体（IL-6R）胞外区真核表达载体，检测其在体外培养细胞中的表达。方法利用PCR扩增IL-6R胞外区，克隆到pcDNA3．1（＋）中，用双酶切、测序鉴定。重组质粒通过脂质体转染HL-60细胞，用G418进行筛选，利用Western印迹检测IL-6R蛋白表达。结果PCR扩增出1218bp的目的片段，双酶切和测序结果显示重组质粒正确。Western印迹结果显示转染细胞能够表达目的蛋白。结论成功构建了人IL-6R胞外区真核表达载体，并且能够在真核细胞中表达。

Construction of Eukaryotic Expression Vector Containing Extracellu- lar Domain of Human IL-6 Receptor and its Expression in vitro

Objective To construct the eukaryotic expression vector encoding extracellular domain of human IL-6 receptor （IL-6R） and examine its expression in transfected cells in vitro. Methods DNA fragment of extracellular domain of human IL-6R was amplified by PCR and cloned into pcDNA3.1 （ ＋ ） vector. The recombinant plasmid was verified by restriction enzyme digestion and DNA sequencing. HL-60 cells were transfected with the recombinant plasmid by means of liposome. The transfectants were screened by G418 selection. Expression of IL-6R protein product in transfected cells was detected by Western blot. Results The target 1 218 bp fragment of extracellular domain of human IL-6R was amplified by PCR. Restriction enzyme digestion and DNA sequencing confirmed the fragment insert in the recombinant plasmid. Western blot showed the expression of IL-6R in transfected cells. Conclusion Eukaryotic expression vector expressing extracellular domain of human IL-6 receptor was successfully constructed and was able to express in eukaryotic cells.