霍乱弧菌O1群稻叶型流行株和非流行株的蛋白质谱特征分析

Proteome analysis of the total protein in epidemic and non-epidemic Vibrio cholerae O1 Serotype Ogawa strains by 2-DE

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 study the differences on proteins between non-epidemic strain and epidemic strain of Vibrio cholerae （V.cholerae） by two dimensional electrophoresis（2-DE） and TOF-TOF-MS. Methods Total protein was extracted from V. cholerae, and separated by 2-DE under immobilized pH gradients（IPG）, and then stained with Coomassie Brilliant Blue, and analyzed by ImageMaster 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 （1 081±16） protein spots,and pI value was mainly localized between 4.0 and 7.2.Matching spots were （1 057±28） in two repeats electrophoregrams, with a matching ratio of 97.85%. Conclusion The different protein spots were successfully identified with high quality and sharpness separation by 2-DE and TOF-TOF-MS, and serve as a valuable resource for proteomics research of V.cholerae.