摘要
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。
The total protein was extracted from Camellia oleifera kernel using different methods, and the conditions of two - dimensional eleetrophoresis were also explored and optimized to establish a set of opti- mal 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 im- proved TCA -acetone method. 816 protein spots were obtained, and most of protein spots mainly concentrated at the isoelectrie 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.
出处
《中国油脂》
CAS
CSCD
北大核心
2018年第2期79-83,共5页
China Oils and Fats
基金
贵州省科技支撑计划项目(黔科合支撑[2016]2517-1
黔科合支撑[2016]2519号和黔科合支撑[2013]201号)
中央高校基本科研业务费重大专项(DC201501070102)
关键词
油茶
种仁总蛋白
提取方法
双向电泳
Camellia oleifera
total kernel protein
extraction method
two -dimensional electrophoresis
