Establishment Of Two Dimensional Gel Electrophoresis System In The Research Of Proteomic Of Vibrio Cholerae

With the accomplishment of Human Genome Project, the research of life science has been entering Post-genome era. Proteomics is a new subject from genomics and mainly studies proteins of cells and its regulation, including characterization of proteins, post-translational modifications and the interation of proteins. Now proteomics research has become one of the hot fields of 21 century life science. As main separation technique, Two dimensional gel electrophoresis can separate tens of thousands proteins.The strain of Vibrio cholerae N16961,JS32,D118,MO45,93284 et al were used to Establish Two dimensional Gel Electrophoresis system.Sonication- Urea-thiourea-CHAPS-DTT was employed to extract whole Vibrio cholerae's proteins.these proteins were applied on to the immobilized pH gradient strips(PH4-7,6-11); the total Vh( the product of voltage and hours) is 56kVh-66kVh . After two steps equilibrium, the first dimension IPG was applied on to the second dimension SDS-PAGE(12.5%, 1.0mm). Resulting gels were stained by commassie brilliant blue. The images of two-dimensional electrophoresis were obtained by ImageScanner optical density scaner. almost 700 spots can be obtained from two-dimensional electrophoresis map.these maps has high resolution and high reproducibilityUsing this2-DE technique, high-quality of whole Vibrio cholerae's proteins could be extracted and satisfactory 2-DE maps with high reproduction and resolution power of protein were obtained,suggesting that the 2-DE technique developed in this study could absolutely meet therequirement of further proteomics study.