Objective To study the different proteins of human and environment Vibrio cholerae (V. cholerae) by two dimensional electrophoresis (2-DE) and TOF-TOF-MS. Methods Total cellular pro-tein was extracted by lysate from V. cholerae, and proteins were separated by 2-DE under immobilized pH gradients(IPG), then electrophoregrams were dealed with coomassie brilliant blue, and analyzed by Im-ageMaster 2D Elite 5.0, finally the different protein spots were identified by TOF-TOF-MS. Results High repetitive 2-DE maps were obtained. 2-DE and image analysis revealed 1032±22 protein spots, and PI value was among 4.00 to 7.20. Matching spots were 1025±24 in two repeats electrophoregrams and matching ratio was 96.30%. Conclusion The different protein spots were successfully established with high quality and sharpness separation by 2-DE and TOF-TOF-MS, which stands as a valuable resource for proteomics re-search of V. cholerae.

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