The 7 strains with antimicrobial activity were screened as samples and verification by PCR amplification has antimicrobial peptide genes of PlnF, PlnE, PlnN, PlnJ, and PlnK. Then designed a primer to introduce Nco I and Xho I sites into both ends of the antibacterial peptide genes with the treated of pET28a plasmid to obtain recombinant plasmid using T4 ligase, and the pET28a-peptide was transformed into E.coli BL21(DE3). The experiment results showed that the inhibition zone of the pET28a-peptide was the PlnF, after 0.5 mmol/L IPTG inducted for 6 h after the OD₆₀₀=0.6 of the E.coli LB. The cell in the 400 W, ultrasonic for 4 s, with intermittent 5 s for crushing, and then with 8 mol/L urea denaturation and renaturation with Ni purified, the PlnF purified protein has the antibacteria effects to the Staphylococcus aureus with (13.43±0.21) mm compare with the control group. Further by Tricine-SDS-PAGE electrophor-esis and nanoLC ESI/MS/MS verification, the homology of the target protein and PlnF antimicrobial peptide with 97.6%.