樟树叶片蛋白提取方法及其应用

Protein extraction method from Cinnamomum camphora leaves and applications in five chemotypes

提取高质量的蛋白质是蛋白质组研究的基础。以樟树5种化学类型(芳樟、异樟、脑樟、油樟和龙脑樟)叶片为材料,采用Tris-HCl结合TCA/丙酮沉淀法提取总蛋白质,并对提取效果进行检测。定量检测结果表明,所有样品的蛋白量都可以满足下一步消化实验的要求;PAGE电泳结果表明,Tris-HCl结合TCA/丙酮沉淀法所得图谱背景清晰,蛋白质信息量很大。检测结果说明用本试验的方法可获得高效、丰富的樟树叶片总蛋白质,能够满足蛋白全谱、磷酸化修饰、目标蛋白分析等相关研究,为深入开展不同化学类型樟树蛋白质组学研究奠定基础。

High quality protein extracting is the foundation for proteomics research. Total protein was extracted for five chemotypes of Cinnamomum camphoraby, by Tris-HCl extraction and TCA/acetone precipitation method, and the extraction effect was evaluated also. The results showed that the amount of protein in all samples could meet the requirements for next digestion experiments through quantitative analysis. It was showed that protein profiles had clear background patterns and more abundant information on PAGE after electrophoresis. The protein extracted by Tris-HCl extraction and TCA/acetone precipitation method could meet the full spectrum of total protein, protein phosphorylation, target protein analysis and other related research to provide a solid foundation for further study in proteome research.