中文摘要：液相色谱-串联质谱联用技术在转基因作物蛋白质检测中的应用越来越广泛。目前，需要研发一个没有蛋白质参考标准品的快速蛋白质检测技术。本文研发了一个检测靶标蛋白的敏感、特异方法，该方法可在没有参考标准或样品富集的情况下，检测转基因玉米叶粗提物的靶标蛋白。作者用混合Q-TRAP质谱仪来监测转基因和非转基因样品中靶标蛋白的所有潜在胰蛋白酶肽，并用MIDAS方法通过Mascot数据库检索来识别初始肽/蛋白质。另外，通过转基因和非转基因样品间的直接比较来进行进一步的识别。研究结果表明，在没有蛋白质参考标准品或抗体的情况下，基于MIDAS的靶向蛋白质组学质谱法是在转基因作物的研究和开发的早期阶段检测新型蛋白质的理想方法。
外文摘要：BACKGROUND：Liquid chromatography coupled with tandem mass spectrometry is increasingly used for protein detection for transgenic crops research. Currently this is achieved with protein reference standards which may take a significant time or efforts to obtain and there is a need for rapid protein detection without protein reference standards.
RESULTSA：sensitive and specific method was developed to detect target proteins in transgenic maize leaf crude extract at concentrations as low as approximate to 30ngmg(-1) dry leaf without the need of reference standards or any sample enrichment. A hybrid Q-TRAP mass spectrometer was used to monitor all potential tryptic peptides of the target proteins in both transgenic and non-transgenic samples. The multiple reaction monitoring-initiated detection and sequencing (MIDAS) approach was used for initial peptide/protein identification via Mascot database search. Further confirmation was achieved by direct comparison between transgenic and non-transgenic samples. Definitive confirmation was provided by running the same experiments of synthetic peptides or protein standards, if available.
CONCLUSIONA：targeted proteomic mass spectrometry method using MIDAS approach is an ideal methodology for detection of new proteins in early stages of transgenic crop research and development when neither protein reference standards nor antibodies are available.
外文关键词：mass spectrometry；target proteomics；protein detection；MIDAS；transgene analysis
作者：Schacherer, Lindsey J；Xie, Weiping；Owens, Michaela A；等
作者单位：DuPont Pioneer Agr Biotechnol
期刊名称：JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE
期刊影响因子：2.076
出版年份：2016
出版刊次：9
点击下载：在没有蛋白质参考标准品的情况下，利用靶向蛋白质组学质谱法快速检测转基因作物中的蛋白质