Abstract: The pcDNA5/FRT-SlSCP-2 and positive control pcDNA5/FRT-GFP eukaryotic expression vector was constructed and transient transfected into CHO cells. The Spodoptera litura sterol carrier protein (SlSCP-2) protein expression in the cells was confirmed by western blotting and fluorescence inverted microscope respectively. After a 24 h transfection of CHO cells with the recombinant plasmids, the positive cell clones expressed SlSCP-2 and GFP correctly. SlSCP-2 can increase the uptake of cholesterol by cells. Meanwhile, the suitable concentration of hygromycin B was determined for the screening of stable transfected cell lines. The CHO cell lines with Eukaryotic expression of SlSCP-2 protein and determination of the screening concentration of Hygromycin B are the basis for the establishment of a stably cell line which was used for insecticide screening.