In this study, we show the expression and purification of the human recombinant farnesoid X receptor (FXR)- ligand binding domain (LBD) protein in E. coli using a double cistronic vector, pACYCDuet-1, as a soluble form. We describe here the expression and characterization of a biologically active FXR-LBD (248-476). When expressed in the influence of bacterial promoters (PT7 and PTac) of the single cistronic expression vectors, the human recombinant FXR-LBD (248-476) was found to be totally insoluble. However, by using a double cistronic expression vector, we were able to obtain the human recombinant FXR-LBD (248-476) in a soluble form. To allow for biological activities, we have subcloned into the pACYCDuet-1 vector, expressed in E. coli cells at some optimized conditions, and purified and characterized the human recombinant active FXR-LBD (248-476) proteins using the fluorescence polarization assay. This suggests that the expression of FXR-LBD in a double cistronic vector improves its solubility and probably assists its correct folding for the biologically active form of the proteins. We suggest that this may represent a new approach to high expression of other nuclear receptors and may be useful as well for other classes of heterodimeric protein partners.