作　　者： ; ; ; ; ; ; ; ; ;

机构地区： 北京林业大学

出　　处： 《东北林业大学学报》 2011年第8期55-57,共3页

摘　　要： 以银杏小孢子叶球为材料,比较了不同的提取的方法、蛋白质裂解液组成、pH值范围以及不同质量浓度SDS-PAGE凝胶对双向电泳的影响。结果表明:TCA/丙酮/酚法比TCA/丙酮法更适合于银杏小孢子叶球蛋白的提取;含有硫脲的蛋白质裂解液Ⅱ,可显著提高蛋白质的溶解性,在2-DE图谱上可分辨出748个蛋白质点,比用蛋白质裂解液Ⅰ提取的蛋白点数多620个。采用pH3-10的胶条分离时,蛋白点集中在pH=4～8,使用pH=4～7的胶条,图谱胶点重叠少,也较清晰,更利于低丰度蛋白的检出。用TCA/丙酮/酚提取蛋白,并以蛋白质裂解液Ⅱ溶解蛋白,采用pH=4～7线性17 cm的IPG胶条1,2%SDS-PAGE凝胶进行双向凝胶电泳,银杏小孢子叶球蛋白质能得到较好的分离,图谱清晰。 The effects of different protein extraction methods, lysis buffers, pH ranges of IPG strip and different gel concentrations on two-dimensional gel clectrophoresis were studied by using male flower buds of Ginkgo biloba as experimental materials. Result shows that, compared TCA-acetonc-phenol protocol with TCA-acetone protocol, the former is more suitable. The protein solubility of the samples dissolved in lysis buffer Ⅱ containing thiourea was greatly enhanced, and about 748 protein spots were detected on 2-DE gels, which was 620 more than that in lysis buffer Ⅰ. More spots were assembled from pH 4 to pH 8. Replaced pH 3-10 strips with pH 4-7 strips, the spots were shown in a clearer way. When the proteins were extracted by the TCA-acetone-phenol method, dissolved with the lysis buffer Ⅱ, with 17 cm, pH 4-7 IPG strip, and 12% concentrations of the SDS-PAGE gel, they were better separated.

关 键 词： 银杏 丙酮 酚 双向电泳技术

领　　域： [农业科学] [农业科学]