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| 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 |
| KeyWord:Camellia oleifera kernel protein two-dimensional gel electrophoresis TCA-acetone precipitation method BPP extraction method proteomics |
| FundProject: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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| Abstract: |
| 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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