Objective: To clone and express the human cytochrome P450 2E1(CYP2E1) gene. Methods: Using RT-PCR, a full length gene encoding CYP2E1 was successfully cloned and its expression was analyzed in the host cell E. coli BL21(DE3)by SDS-PAGE. Results: Except a C→T mutation at 1 263 nucleotide site, the nucleotide sequences of this CYP2E1 gene are consistent with the sequences in GenBank. The CYP2E1 gene was successfully inserted into the expression vector pET-32a (+) and expressed in the host cell E. coli BL21(DE3). The CYP2E1 protein with relative molecular mass of approximately 67ku was observed on the SDS-PAGE gel. Conclusion:The cloning and expression of CYP2E1 provide the foundation for following pre-clinical drug safety evaluation and the analysis of drug metabolism.