白桦花芽蛋白质双向电泳技术的建立

Proteome Two-Dimensional Electrophoresis of Flower-Bud of Betula platyphylla

目的:初步建立白桦花芽蛋白质组研究的双向电泳(2-DE)技术,以提高其分辨率及重复性。方法:以白桦花芽为实验材料,对关键环节如样品处理、上样量和染色方法等进行一系列优化。结果:蛋白上样量为50～60μg、银染剂中采用0.16%的AgNO3染色12min,即可获得蛋白点的形态规则、稳定性高、重复性好的2-DE图谱。结论:建立了适合多糖、醌类等次生代谢物质含量较多的白桦花芽蛋白提取方法,为开展白桦发育生物学研究奠定了基础。

Objective： To establish two-dimensional electrophoresis（2-DE） patterns of flower-bud for Betula platyphylla. Methods： A series of important factors, such as sample handling, staining methods, loading amount and so on were optimized. Results： On of condition of 50-60 μg sample of loading amount, 0.16% of AgNO3 concentration and 12 min of silver staining, electrophoresis map in high resolution was obtained. Conclusion： The technique of protein extraction which is suitable for B.platyphylla containing some secondary metabolites has been established. A scientific basis for molecular phylogeny of B.platyphylla is provided.