Construction of eukaryotic expression vector containing human sperm protein 17 gene and its expression in ovarian carcinoma cells

Objective To construct eukaryotic expression vector of pcDNA3.1(+)/hSp17 containing human sperm protein 17(hSp17) cDNA,so to provide basis for its clinical application.Methods hSp17 cDNA was obtained by PCR,and to construct eukaryotic expression vector pcDNA3.1(+)/GNLY by NheⅠ,KpnⅠdigestion and sticky end ligation techniques.The recombinant plasmid was transfected into H08910 cells with lipofectamine 2000.Expression of hSp17 in the transfected cells was observed by immunocytochemistry.Cells with stable expression of hSp17 were selected by G418.Results Sequencing and restriction enzyme digestion showed pcDNA3.1(+)/GNLY was correctly constructed.The stably hSp17 gene transfected HO8910 cell clones were obtained through G418 screening and immunocytochemistry assay were further performed to confirm the expression of hSp17 in the HO8910 cells.Conclusion An ovarian carcinoma cell model that stably expressed hSp17 can be successfully established;which can provide basis for immunotherapy of ovarian carcinoma.