油茶种仁总蛋白提取及双向电泳分析

以油茶种仁为实验材料,用不同方法提取油茶种仁总蛋白,同时对双向电泳条件进行了探索与优化,建立了一套适合油茶种仁总蛋白双向电泳分析的优化体系。结果表明:改良后的TCA-丙酮法可以有效提取出油茶种仁总蛋白,得到了816个蛋白点,蛋白点主要集中在等电点p H 4.0~6.0之间;选用17 cm p H 4~7 IPG胶条,优化的等电聚焦程序为250 V 0.5 h、1 000 V 2 h、8 000 V5 h、8 000 V 60 000 Vh和500 V 24 h,可获得背景清晰、分辨率较高的双向电泳图谱。

Extraction of total protein from Camellia oleifera kernel and two-dimensional electrophoresis analysis

The total protein was extracted from Camellia oleifera kernel using different methods, and the conditions of two-dimensional electrophoresis were also explored and optimized to establish a set of optimal system of two-dimensional electrophoresis analysis for total protein in Camellia oleifera kernel. The results showed that total protein could be effectively extracted from Camellia oleifera kernel by the improved TCA-acetone method. 816 protein spots were obtained, and most of protein spots mainly concentrated at the isoelectric points pH 4.0-6.0. The optimal two-dimensional electrophoresis system was obtained as follows: length of IPG strip 17 cm with pH 4-7,isoelectric focusing program 250 V 0.5 h, 1 000 V 2 h, 8 000 V 5 h, 8 000 V 60 000 Vh and 500 V 24 h. Under these conditions, a 2-DE map with clear background and higher resolution could be obtained.