橡胶树大小橡胶粒子的差速离心分离及其差异表达蛋白质的研究

Studies on the Separation and the Differentially Expressed Proteins of the Large and the Small Rubber Particles of Rubber Trees(Hevea brasiliensis Muell.Arg.)

为了鉴定巴西橡胶树胶乳中大小橡胶粒子之间的差异表达蛋白质以及揭示橡胶生物合成的分子机制,采用差速离心的方法,从橡胶树的新鲜胶乳中分离纯化总橡胶粒子、大橡胶粒子和小橡胶粒子,用扫描电镜和SDS-PAGE以及质谱等方法分别检测和鉴定大小橡胶粒子的分离效果及其差异表达蛋白。结果表明,大小橡胶粒子可用差速离心法得到有效的分离和纯化,它们在SDS-PAGE电泳图谱上表现出差异表达的橡胶粒子蛋白,如分子量为22.0kDa、24.5kDa和120kDa蛋白等,并对其中分子量为24.5kDa的小橡胶粒子蛋白(SRPP)进行了质谱分析。橡胶树大小橡胶粒子差速离心法的建立,为大小橡胶粒子差异表达蛋白质的分离和鉴定奠定了一定的基础,将进一步促进橡胶树橡胶粒子的起源和发育,以及橡胶生物合成分子机制的研究。

In order to identify the differential expressed proteins between the small rubber particles （SRPs） and the large rubber particles （LRPs） in the latex of Hevea brasiliensis （rubber tree） and to unravel the molecular meehanisms of rubber biosynthesis, SRPs, LRPs and the total rubber particles were completely separated and purified from the fresh latex using differential centrifugation, and further detected through scanning electron microscope （SEM）, respectively. One-dimensional SDS-PAGE analysis demonstrated that the protein profile of the LRPs was different with that of the SRPs, including the differentially expressed proteins with appropriate mass weight of 22.0 kDa, 24.5 kDa and 120 kDa, and so on. One protein expressed only on the SRPs, SRPP, was further identified using nano-LC MS/MS. The separation of LRPs and SRPs from the latex would facilitate both the identification of the differential expressed proteins between LRPs and SRPs, and the further investigation into the molecular mechanisms not only promote the biogenesis and the development, but also promote the rubber biosynthesis of the rubber particle of rubber trees.