The coding region of FoxA1，amplified by PCR, was cloned to construct the lentivirus vector plv-rFoxA1-IRES-EGFP. The recombinant plasmid was transfected transiently into 293T cells and the expressions of GFP and FoxA1 were confirmed by fluorescent microscopy and Western blot. The plv-rFoxA1-IRES-EGFP was cotransfected with the packaging plasmids △8.91, pVSV-G to 293T cells to produce the lentivirus. The results show that lentiviral vector that expresses rat FoxA1 was successfully constructed, and infection ability of the recombinant lentivirus was confirmed by the effective expression of FoxA1 protein in cells. These results provided an important material to study the FoxA1 functions in the future.