霍乱弧菌菌体蛋白双向电泳技术的建立  

Proteome Analysis of the Total Protein in Vibrio cholerae by 2-DE

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

机构地区：[1]广州市疾病预防控制中心,广州510080

出　　处：《热带医学杂志》2008年第4期335-337,共3页Journal of Tropical Medicine

基　　金：广州市卫生局重点专科项目(No.2006-zdi-11)。

摘　　要：目的建立霍乱弧菌全菌体蛋白的双向电泳技术,获得分辨度高、重复性好的双向电泳图谱。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的双向电泳图谱,并利用ImageMaster 2D Elite5.0图象分析软件进行分析,所得的数据用SPSS15.0进行统计分析。结果得到了(1081±16)个蛋白斑点,蛋白主要集中在pI4.24～7.20之间,重复胶的匹配点数为(1057±28),匹配率为97.85%。结论建立了霍乱弧菌全菌双向电泳分析方法,2-DE图谱中蛋白位点的分辨率和重复性非常高,获得了较为理想、清晰的双向电泳图谱,为进一步研究其蛋白质组学奠定了基础。Objective To establish of two dimensional electrophoresis （2-DE） for proteome analysis of Vibrio cholerae（V.cholerae）. Method Total cellular protein was extracted from V.cholerae, and proteins were separated by 2-DE under immobilized pH gradients （IPG）,then electrophoregrams were obtained by Coomassie brilliant blue staining, and analyzed by ImageMaster 2D Elite 5.0. Result High quality and reproducible 2-DE maps were obtained, 2-DE and image analysis revealed （1081±16） protein spots, and pI value was mainly ranged from 4.24 to 7.20. The matching spots were （1057±28） in two repeat electrophoregrams, with matching rate of 97.85%. Conclusion 2-DE of V.cholerae has been successfully established with high quality and resolution, which is valuable for proteomics research of V.cholerae.

关 键 词：霍乱弧菌 双向电泳 蛋白质组学 

分 类 号：R378.3[医药卫生—病原生物学]