This research amplified the phytase gene of Aspergillus niger F246 by the polymerase chain reaction(PCR), which is selected and identified by ourselves.Then the product of PCR was inserted respectively into the lactobacillus expression vector pMG36e to construct the new type of expression vector pMG36e-phyA. The genomine DNA of the Aspergillus niger F246 strain is used as the template for PCR,and the PCR product was subcloned into pMD18T. After sequncing the coding region and analying the sequnce of amino acids, the product of PCR was cutted from pMD18T and subcloned into pMG36e for the engineering strain of Lactobacillus MG1363-pMG36e-phyA. The construction of new recombinant expression vector pMG36e-phyA for the phyA gene lays the base of the study on obtaining large and high active phytase and developing the new microbial ecologicalagent.