霍乱弧菌O1群有毒株和无毒株的蛋白质谱特征分析  

Proteome analysis of the total protein in Vibrio cholerae by 2-DE

作　　者：陈守义[1] 刘俊华[1] 张欣强[1] 胡玉山[1] 李钏华[1] 王鸣[1] 

机构地区：[1]广州市疾病预防控制中心,510080

出　　处：《中华微生物学和免疫学杂志》2008年第11期963-966,共4页Chinese Journal of Microbiology and Immunology

基　　金：广州市卫生局重点专科项目（2006-zdi-11）

摘　　要：目的利用双向电泳和质谱鉴定技术探讨环境和人体中霍乱弧菌蛋白表达的差异。方法利用适当的裂解液处理霍乱弧菌，提取全菌蛋白；采用pH梯度等电聚焦对全菌蛋白进行双向电泳；考马斯亮蓝染色后获得双向电泳图谱，并利用ImageMaster 2D Elite5．0图像分析软件进行分析，所得的数据采用SPSS15．0进行统计分析，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1032±22个蛋白斑点，蛋白主要集中在等电点（PI）4．00—7．20之间，重复胶的匹配点数为1025±24，匹配率为96．30％；发现了有明显差异的21个蛋白点，并用质谱鉴定了其中4种蛋白。结论获得了环境和人体中霍乱弧菌蛋白的差异表达蛋白。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 protein 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 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 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 research of V. cholerae.

关 键 词：霍乱弧菌 双向电泳 基质辅助激光解吸-电离飞行时间质谱 蛋白质组学 

分 类 号：R378.3[医药卫生—病原生物学]