龙眼胚性培养物高分辨率蛋白质双向电泳技术

High-resolution two-dimensional electrophoresis technology for the analysis of the proteins of embryogenic cultures in longan(Dimocar pus longan Lour.)

试验以龙眼胚性培养物为材料,通过对小型垂直板双向电泳和固相pH梯度凝胶双向电泳比较分析,进行龙眼胚性培养物高分辨率蛋白质双向电泳技术方法的研究.结果表明,采用固相pH梯度凝胶双向电泳可以极大地提高分辨率,相同龙眼胚性培养物的蛋白样品染色后所得的蛋白质谱点数从334左右增加到810左右;由于龙眼胚性培养物的蛋白等电点集中在3.5-7.0之间,采用13 cm的IPG胶条时,以pH 4-7的IPG胶条分离蛋白的效果佳.

In this experiment the high-resolution two-dimensional electrophoresis technology for the analysis of the proteins of embryogenic cultures in longan was developed by the comparison between the mini two-dimensional vertical electrophoresis and the immobilized pH gradients two-dimensional electrophoresis. The results showed that the resolution was improved by the immobilized pH gradients two-dimensional electrophoresis, which obtained 810 points using the same embryogenesis materials, while 334 points were obtained by the mini two-dimensional vertical electrophoresis. The best result was obtained under the condition of the pH value between 4 and 7 in the IPG of the 13 cm length, because the isoelectric points of the proteins in longan embryogenic cultures were focused on the pH value between 3.5 and 7.