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| 海南油茶种仁蛋白质提取及双向凝胶电泳-质谱初步鉴定 |
| Extraction of kernel protein from Camellia oleifera in Hainan and preliminary identification by two-dimensional gel electrophoresis combined with mass spectrometry |
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| DOI：10.12166/j.zgyz.1003-7969/2020.03.021 |
| 中文关键词: 油茶 种仁 蛋白 双向凝胶电泳 TCA-丙酮沉淀法 BPP抽提法 蛋白质组学 |
| 英文关键词:Camellia oleifera kernel protein two-dimensional gel electrophoresis TCA-acetone precipitation method BPP extraction method proteomics |
| 基金项目:海南省重大科技计划项目（ZDKJ2017004）；海南省重点研发专项（ZDYF2016157） |
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| 中文摘要: |
| 以海南油茶成熟种仁为试验材料，比较2种不同方法对该植物蛋白提取效率的影响，同时结合单向SDS-聚丙烯酰胺凝胶电泳（SDS-PAGE）、双向凝胶电泳技术（2-DE）、质谱（MS）鉴定手段和Mascot Distiller软件进行分析。结果表明：改良后的BPP抽提法可以有效提取油茶种仁蛋白，通过Image Master 2D Platinum 5.0分析后，在凝胶图上共检测出847个蛋白点，集中分布在等电点（pI）pH 5.0~6.0、相对分子质量86~42 kDa范围内；其图谱背景清晰，蛋白点形状圆润、分辨率高、分布均匀，基本无明显拖尾现象；选取12个高分辨率的蛋白点，经质谱成功鉴定了10个，分别代表10种蛋白质，鉴定成功率达83.3%。BPP抽提法适用于后续油茶种仁的蛋白质组学分析研究。 |
| 英文摘要: |
| The protein was extracted from mature kernel of Camellia oleifera in Hainan using two different methods, and analyzed by one-dimensional SDS-PAGE, two-dimensional gel electrophoresis (2-DE), mass spectrometry (MS) and Mascot Distiller software. The results showed that the protein could be effectively extracted from Camellia oleifera kernel by the improved BPP extraction method. Approximately 847 protein spots were detected in the 2-DE gel image analyzed with Image Master 2D Platinum 5.0, and most of the protein spots mainly concentrated at the range of isoelectric points pH 5.0-60 and relative molecular weights (Mr) 86-42 kDa. The background of gel image was clear, and the protein spots had regular shape, high resolution and uniform distribution, and there was no obvious tailing phenomenon. Twelve high resolution protein spots were selected, and ten protein spots represented ten unique proteins were successfully identified by MS, and the identification success rate was up to 833%. The BPP extraction method could be applied in the further study of Camellia oleifera kernel proteomics. |
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