Aim Trying to establish a lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) bicistronic gene expression vector and detect its expression and function in 293T cell, to explore the role of LOX-1 in atherosclerosis and lay a foundation for establishing intervention mechanism of LOX-1 targets. Methods First human LOX-1 gene was obtained from human cDNA by PCR and cloned into T plasmid. After sequencing, recombinant T plasmid was subcloned into bicistronic eukaryotic expression plasmid pIRES2 AcGFP1-Nuc. Next the bicistronic recombinant expression plasmid was transfected into 293T cells by liposome. Plasmid transfection efficiency was detected by inverted fluorescence microscope. Then the expression of exogenous human LOX-1 gene and protein in 293T cells was detected by RT-PCR and Western blot. The expression of human LOX-1 in 293T cells membrane was detected by confocal laser. Finally LOX-1 functions of binding with ox-LDL in 293T cell membrane was detected by confocal laser and flow cytometry. Results The pIRES2-LOX-1 bicistronic recombinant plasmid was constructed successfully. After transfection, green fluorescent protein was detected and the abundant LOX-1 mRNA and protein were expressed in the transfected cells, moreover human LOX-1 were expressed in the 293T cells membrane and could bind with ox-LDL. Conclusion We have successfully established a LOX-1 gene bicistronic expression vector and on the basis we testified human LOX-1 expressed in 293T cells membrane and could bind with ox-LDL, which layed a foundation for further study of its effect in atherosclerosis and establishing intervention mechanism of LOX-1 targets.