Abstract: The secreted proteins of Vibrio anguillarum MN,V.anguillarum-3101 and Vibrio fischeri which were isolated and conserved by our laboratory,were extracted.The components of secreted proteins were separated by SDS-PAGE.The 5 high expressed protein bands in the SDS-PAGE were identified by mass spectrometry MALDI-TOF/TOF as the metalloprotease of V.anguillarum-MN,the metalloprotease of V.anguillarum-3101,the zinc metalloproteinase of V.anguillarum-3101,the hypothetical protein VFMJ11_1094 and outer membrane protein of V.fischeri-ES114.After downloading the sequences of above proteins from NCBI database,PCR primers were designed and specific DNA bands were amplified,cloned and sequence analysed.5 signal peptides were given by using the online software SignaIP 3.0,namely angMN-35,ang3101-35,ang3101-25,vf-38 and vf-23.The cellular were localization of the secreted sequences were analyzed by PSORT.And we found that all the signal peptides located in the outer membrane or periplasmic space of the cell.The results may be useful for construction of a secreted vector.