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目的探讨霍乱弧菌O1群稻叶型流行株和非流行株的蛋白表达的差异。方法利用裂解液处理霍乱弧菌,提取全菌蛋白;采用pH梯度等电聚焦对菌蛋白进行双向电泳;考马斯亮蓝染色后获得的双向电泳图谱,并利用ImageMaster2DElite5.0图象分析软件进行分析,找出差异蛋白;在此基础上,胰蛋白酶消化这些特殊差异蛋白,并进行质谱(MALDI-TOF-MS)分析。结果获得了(1081±16)个蛋白斑点,蛋白主要集中在pI4.00～7.20之间,重复胶的匹配点数为(1057±28),匹配率为97.85%;发现了有明显差异的21个蛋白点,并进行了质谱鉴定。结论获得了霍乱弧菌O1群稻叶型流行株和非流行株的差异表达蛋白。
Objective To investigate the differences in the protein expression between the Vibrio cholerae O1 group and the non-epidemic strains. Methods Vibrio cholerae was treated with lysate and the whole bacterial protein was extracted. Two-dimensional gel electrophoresis (PAGE) was performed using pH gradient isoelectric focusing. The two-dimensional electrophoresis pattern was obtained after staining with Coomassie Brilliant Blue and analyzed by ImageMaster2DElite5.0 image analysis software. On the basis of this, trypsin digestion of these special differential proteins, and analysis by mass spectrometry (MALDI-TOF-MS). RESULTS: A total of 1081 ± 16 protein spots were obtained. The protein concentration was mainly between pI4.00 and 7.20, and the number of matched spots was (1057 ± 28) and the matching rate was 97.85%. There were 21 significant differences Protein spots, and identified by mass spectrometry. Conclusion The differentially expressed proteins of Vibrio cholerae O1 group rice leaf-type and non-epidemic strains were obtained.