【摘要】 橡胶粒子是巴西橡胶树(Hevea brasiliensis)乳管细胞内进行橡胶生物合成的亚细胞结构;对橡胶粒子的蛋白质组学研究,可为揭示天然橡胶生物合成机理提供线索。采用5种提取液分别提取橡胶粒子蛋白,并对橡胶粒子蛋白进行SDS-PAGE和16-BAC/SDS-PAGE电泳分离及MALDI TOF/TOF串联质谱分析,证明不同提取液抽提的橡胶粒子蛋白具有不同组成,发现分子量较高的橡胶延伸因子家族蛋白更难从橡胶粒子上被洗脱和提取。通过检索橡胶树转录组数据库,鉴定了3个新的橡胶粒子蛋白,即醌氧化还原酶、含蓖麻毒素B链凝集素结构域蛋白及枯萎/脱水相关蛋白。本研究建立的橡胶粒子蛋白质提取和分离方法,为进一步鉴定低丰度和具有重要功能的橡胶粒子蛋白提供了参照体系。更多还原

【Abstract】 Rubber particles are organelles responsible for the biosynthesis of natural rubber in Hevea brasiliensis(rubber tree).Proteomics investigations into rubber particles may provide clues for elucidating the mechanisms of natural rubber biosynthesis.In this study total proteins extracted from rubber particles using five different buffers were subjected to SDS-PAGE and 16-BAC/SDS-PAGE analysis,respectively,and then analyzed by MALDI TOF/TOF tandem mass spectrometry.The data demonstrated that rubber particle proteins extracted with different buffers had different components,and proteins with higher molecular weight in the rubber elongation factor family were not easily extracted from the rubber particles.Three novel rubber particle proteins,quinone oxidoreductase,protein containing ricin B chain lectin domain,and senescence/dehydrationrelated protein,were identified through searching a transcriptome database of H.brasiliensis.Our study establishes a reference system which is helpful to identify novel rubber particle proteins with low-abundance and important biological functions.更多还原