Construction and identification of eukaryotic expression vector of mouse CD25 extracellular domain

Objective To construct a eukaryotic expression vector of mouse CD25 extracellular domain and to express it in CHO. Methods The prokaryotic expression vector PET-32a-CD25e was digested with EcoR I and Hind III. The target sequence was subcloned into eukaryotic expression vector pcDNA3.1/Myc-his-(-). The recombinant plasmid was identified through enzyme cleaving and sequencing before expression. The recombinant plasmids were transferred into CHO by Lipofectamine 2000. The concentration of target molecule in supernatant was detected by ELISA assay. Results The recombinant plasmid pcDNA3.1-CD25 extracellular domain gene was identified by sequencing and could express CD25e protein in CHOcells. Conclusions The mouse CD25 extracellular domain has been successfully expressed in eukaryotic cells, which provided the foundation for further relative studies.