Badri Amine M, Coenen Karine, Vaillancourt Louis-Philippe, Goulet Charles, Michaud Dominique
Département de phytologie, Université Laval, Québec, Canada.
Methods Mol Biol. 2009;483:313-24. doi: 10.1007/978-1-59745-407-0_18.
This chapter presents a general procedure for the on-chip detection and quantitation of low-molecular-weight recombinant proteins in transgenic plant crude extracts by surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF MS). A protocol is first described to detect the protein of interest in crude protein extracts of transgenic plant lines, by differential protein mapping against similar extracts from a control, nontransgenic line. A complementary protocol is then presented to generate a standard curve with the SELDI system, allowing the protein to be quantified in different transgenic lines. Overall, this procedure may be carried out within a few hours, without the need for prior purification or enrichment of the recombinant protein.
本章介绍了一种通过表面增强激光解吸电离飞行时间质谱(SELDI-TOF MS)对转基因植物粗提物中的低分子量重组蛋白进行芯片检测和定量的通用程序。首先描述了一种方案,通过与对照非转基因品系的类似提取物进行差异蛋白质图谱分析,来检测转基因植物品系粗蛋白提取物中的目标蛋白。然后介绍了一种补充方案,用于使用SELDI系统生成标准曲线,从而能够对不同转基因品系中的蛋白质进行定量。总体而言,该程序可在数小时内完成,无需事先对重组蛋白进行纯化或富集。