Proteomic Analysis Of Vibrio Alginolyticus In Response To Four Types Of Antibiotics

Over the past decade,there had been a rapid development of our aquaculture industry. At the same time, various fish diseases were emerging, with the development and the harmfulness was growing. Vibrio is a potential human pathogen; Vibrio alginolyticus is one of the most harmful pathogens which occasionally cause life-threatening infections in Individuals and results in great losses in marine aquacultures of crustaceans and fish. Since wide-ranging outbreak of Vibrio diseases, a large number of antibiotics were used to inhibit fish diseases, but with the widespread use of antibiotics, the drug resistance of Vibrio caused serious losses to aquaculture.In the present study, in order to get the differentially expressed Protein profiles of V. alginolyticus induced by tetracycline, erythromycin, spectinomycin and gentamicin , the whole cell protein of V. alginolyticus was investigated by 2-D SDS-PAGE, and we identified differentially expressed protein spots by mass spectrometry identification. The result of protein profile showed that there was significant difference on the whole cell protein of V. alginolyticus after induced by the four types of antibiotics. There were 13 spots different after analyzed by the software of Image Master 2D Platinum. The result of mass spectrometry identification was as follows:1. There were 3 expressed spots of V. alginolyticus induced by tetracycline. The 3 protein spots were Glycerol-3-phosphate-binding periplasmic protein and Putative 3-keto thiolase Phosphate ABC transporter and periplasmic phosphate-binding protein. Glycerol-3-phosphate-binding periplasmic protein and Putative 3-keto thiolase Phosphate ABC transporter were showed to be up-regulated and periplasmic phosphate-binding protein down-regulated. The result suggested that the antibiotic-resistant mechanism of V. alginolyticus was associated with binding periplasmic protein and transporter protein we also obtained that tetracycline maybe inhibited the synthesis of protein of V. alginolyticus.2. There were 3 spots differentially expressed of V. alginolyticus induced by erythromycin. The 3 protein pots are triosephosphate isomerase and outer membrane pro- tein N and phosphoenolpyruvate carboxykinase. The 3 protein spots were all down- regulated in the result of mass spectrometry identification. The result suggested that the antibiotic-resistant mechanism of V. alginolyticus was associated to outer membrane protein and energy metabolism of V. alginolyticus.3. There were 3 spots differentially expressed of V. alginolyticus induced by spectino- mycin. The 3 protein pots are hypothetical protein VPA1203 and 50S ribosomal protein L10 and ABC-type Fe3+ transport system, periplasmic component. These 3 protein spots were all up- regulated. The result suggested that the antibiotic-resistant mechanism of V. alginolyticus was associated with ribosomal protein and transport system.4. There were 4 spots differentially expressed of V. alginolyticus induced by gentamicin. The 3 protein pots are hypothetical protein VPA1203 and putative histidine- kinase and glucose-specific PTS system component, in addition, there was another protein unknown. Only the unknown protein was expressed up-regulated while the other 3 protein spots were all down- regulated.The four types of antibiotics (tetracycline and erythromycin and spectinomycin and gentamicin) were selected optimally in the study, and the antibiotic-resistant mechanism of V. alginolyticus was discussed from different perspectives. From differentially expressed protein spots, antibiotic-resistant mechanism of V. alginolyticus of which in activator or inactive enzyme were produced, target proteins were modified, efflux pump systems and permeability of outer membrane were changed could be known.