Abstract: For expressing the recombinant GST-FloR1 protein in prokaryote expression system, floR gene was cloned into pGEX-4T-2， and BL21(DE3) codon plus was chose as corresponding expression hosts. The floR1 gene could be successfully expressed in the systems， by optimizing the IPTG induction concentration， incubation temperature and time. The method of tritonurine with sonication lysis of cells was chose to get the relatively good washing effect. After the protein refolding of denaturant inclusion body following dialysis， we got the pure recombinant GST-FloR1 protein by GST affinity columns. A murine anti-FloR antibody was produced by using the recombinant protein of GST-FloR1 as antigen. The antibody lays the foundation for further research on location of FloR protein and inhibition of drug efllux function.