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作 者 ：杨梅, 陈伟, 林思祖, 曹光球

期 刊 ：热带亚热带植物学报 2007年 15卷 5期 页码：438~442

关键词：杉木；叶片；双向电泳；蛋白质组；

Keywords:Chinese fir, Leaves, Two-dimensional polyareylamide gel electrophoresis, Proteome,

摘 要 ：为建立适用于杉木(Cunninghaimia lanceolata)叶片蛋白质组研究的双向电泳技术，对杉木叶片蛋白质的溶解方法、上样量、IEF及SDS－PAGE电泳等关键步骤进行了优化。结果表明，杉木叶片蛋白质主要分布在pH4-7范围；裂解液中含有硫脲（2mmol/L）才能较充分地溶解蛋白，DTT浓度60 mmol/L、上样量1.5mg时得到的图谱分辨率较好且蛋白斑点分布均匀、清晰，拖尾现象明显减少，平衡液Ⅱ中碘代乙酰胺450 mg(15ml)^-1时能提高图谱分辨率；采用与质谱兼容的考马斯亮兰进行染色，得到近700个蛋白点。

Abstract:A two-dimensional polyareylamide gel electrophoresis(2D-PAGE) system was established for proteomic study of Chinese fir (Cunninghaimia lanceolata) leaves. The result shows that the system containing lysis buffer with 2 mmol/L thiourea for complete dissolution of proteins, DTT 60 mmol/L, loaded proteins 1.5 mg for good protein patterns, equilibration liquid with 450 mg(15 ml)^-1 iodoacetamide for high resolution patterns, colloidal coomassie blue compatible with mass spectrometer. More than 700 protein spots with pH 4-7 were detected.

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