2D-DIGE System for Proteomics

蛋白质组学 2D-DIGE 系统

• 批准号: 6877543
• 负责人: RUTH Hogue ANGELETTI
• 金额: $ 18.01万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6877543
• 关键词: biomedical equipment purchase; molecular /cellular imaging; proteomics; two dimensional gel electrophoresis

DESCRIPTION (provided by applicant): Funds are requested for a two dimensional difference gel electrophoresis (2D-DIGE) sample preparation system for proteomics, consisting of devices for: separation, fluorescence imaging, computerized data analysis, robotic spot-picking, proteolytic digestion, and delivery to targets suitable for mass spectrometry analysis. This system provides a semi-quantitative assessment of differential protein expression with enhanced reproducibility between pairs of samples, since fluorescent dyes of different emission spectra are utilized to label the samples. As a result, statistical reliability is increased, and precious samples are more efficiently used. This feature is even more critical for proteomics analysis of samples prepared using the cell sorter or of laser captured cell populations, as described in some projects. The imaging device also permits comparison of protein patterns with potentially phosphorylated proteins detected using a fluorescent dye and with immunoreactive proteins. Investigators participating in this proposal have proteomics applications in signal transduction, cell communications, obesity, diabetes and cancer research and biological discovery. This instrument system will also be useful for testing feasibility of new projects not included in this proposal. The instruments will be operated and supervised by experienced protein chemists and mass spectrometrists within an existing shared protein core facility with a record of productivity and collaborative interactions.

描述（由申请人提供）：申请资金用于蛋白质组学的二维差异凝胶电泳（2D-DIGE）样品制备系统，该系统由以下设备组成：分离、荧光成像、计算机化数据分析、机器人点选、蛋白水解消化和递送至适用于质谱分析的目标。该系统提供了差异蛋白质表达的半定量评估，具有样品对之间增强的再现性，因为不同发射光谱的荧光染料被用于标记样品。因此，统计可靠性得到提高，宝贵的样本得到更有效的利用。如某些项目中所述，该特征对于使用细胞分选仪制备的样品或激光捕获的细胞群的蛋白质组学分析甚至更为关键。成像装置还允许将蛋白质模式与使用荧光染料检测的潜在磷酸化蛋白质和免疫反应性蛋白质进行比较。参与这项提案的研究人员将蛋白质组学应用于信号转导、细胞通讯、肥胖、糖尿病和癌症研究以及生物发现。该仪器系统还将有助于测试本提案中未包括的新项目的可行性。这些仪器将由经验丰富的蛋白质化学家和质谱仪专家在现有的共享蛋白质核心设施内操作和监督，该设施具有生产力和协作互动的记录。

项目成果

The Phage-Encoded N-Acetyltransferase Rac Mediates Inactivation of Pseudomonas aeruginosa Transcription by Cleavage of the RNA Polymerase Alpha Subunit.

• DOI: 10.3390/v12090976
• 发表时间: 2020-09-02
• 期刊: Viruses
• 作者: Ceyssens PJ;De Smet J;Wagemans J;Akulenko N;Klimuk E;Hedge S;Voet M;Hendrix H;Paeshuyse J;Landuyt B;Xu H;Blanchard J;Severinov K;Lavigne R
• 通讯作者: Lavigne R