【摘要】 目的优化双向凝胶电泳的条件,建立适合广州管圆线虫Ⅴ期幼虫的最适二维电泳模型。方法经研磨、反复冻融提取广州管圆线虫Ⅴ期幼虫蛋白质,测定其浓度后分别用pH3～10和pH4～7的固相胶条进行双向凝胶电泳,同时对双向电泳的条件进行调节和优化,银染后进行凝胶图像比较。结果采用pH3～10的胶条分离蛋白时,蛋白质点分布较集中,各蛋白点间间距较小;pH4～7的胶条分辨率较高,蛋白分布均一,获得719个蛋白点,各蛋白点间距相对较大,避免了高丰度掩盖低丰度蛋白点,在相同的pH范围内,检测到的蛋白点更多,灵敏度更高。结论建立和优化适合广州管圆线虫的二维电泳模型,其重复性好、分辨率高,为广州管圆线虫的蛋白质组学研究奠定了基础。更多还原

【Abstract】 To establish and optimize two-dimensional gel electrophoresis(2-DE) models for proteomic analysis of A.cantonensis larvaeⅤ,the protein of larvae Ⅴ of A.cantonensis was extracted by grinding and repeated freezing and thawing,then 2-DE was carried out respectively with the strip of pH3-10 and pH4-7 after measuring the concentration.The gel electrophoresis image was compared after silver staining.The results indicated that the protein spots were more concentrated,and the space among spots were smaller with IPG strip of pH3-10.Then we obtained high-resolute,uniform distributed protein spots,719 protein spots with IPG strip of pH4-7,and the space among spots were relatively large,avoiding to protein spots of high abundance cover the low abundance,it has higher sensitivity.As a result,establishment and optimization of 2-DE models for A.cantonensis was successfully conducted,and 2-DE gel with high resolution was obtained,which might provide foundation for the proteomic analysis of A.cantonensis.更多还原