| 吴波，阮成江，杜维，等.油茶种仁总蛋白提取及双向电泳分析[J].中国油脂,2018,43(2):.[WU Bo, RUAN Chengjiang, DU Wei,etc.Extraction of total protein from Camellia oleifera kernel and two-dimensional electrophoresis analysis[J].China Oils and Fats,2018,43(2):.] |
| 油茶种仁总蛋白提取及双向电泳分析 |
| Extraction of total protein from Camellia oleifera kernel and two-dimensional electrophoresis analysis |
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| DOI： |
| 中文关键词: 油茶 种仁总蛋白 提取方法 双向电泳 |
| 英文关键词:Camellia oleifera total kernel protein extraction method two-dimensional electrophoresis |
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| 摘要点击次数: 1154 |
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| 中文摘要: |
| 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pH 4.0~6.0之间；选用17 cm pH 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 V 5 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。 |
| 英文摘要: |
| 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. |
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