DIFFERENTIAL GEL ELECTROPHORESIS (DIGE) SYSTEM: INFECTIOUS DISEASE

该子项目是利用由NIH/NCRR资助的共享仪器赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。列出的机构是用于资助的，不一定是研究者的机构。描述（由申请人提供）：资金要求为凯克实验室的二维差异（荧光）凝胶电泳（DIGE）系统。该租赁系统与现有的用于蛋白质鉴定的nano-LC-ESI-QTOF一起非常成功地用于鉴定差异表达的蛋白质/生物标志物。DIGE是Keck实验室中最具成本效益、功能强大且高效的蛋白质分析技术，它被大量用于分析来自全细胞、部分纯化的蛋白质提取物和血清等生物液体的各种实验与对照样本。我们的DIGE系统已被用于为19个机构的48名耶鲁大学和32名非耶鲁大学的研究人员进行超过350次定量分析。自2003年12月签订租约以来，对DIGE的需求持续增加。所要求的更新版本的图像分析软件和我们新安装的高通量和灵敏的MALDI TOF/TOF质谱仪所做的改进将使这项技术达到最先进水平，这些改进将进一步增加研究自闭症，药物滥用，脑瘫，精神发育迟滞，早产，哮喘，I型糖尿病，肾脏疾病，肿瘤细胞抗性、髓性白血病的发生、分子骨发育和狼疮。如果不授予这笔赠款，在未来一年内，每月租赁付款增加4倍（因为租赁从2006年5月开始从只支付利息到偿还利息+资本设备成本）将导致DIGE服务费大幅增加。这种增长将对耶鲁和非耶鲁科学界使用这种先进技术产生非常负面的影响。该应用程序的优势包括共同PI监督大型，多研究者NIH合同和中心的经验;主要研究者和其他关键人员的补充生物技术，统计和数据库专业知识;凯克实验室已经在使用的广泛的蛋白质分析技术，以及由几个蛋白质组学和高性能计算中心支持的强大基础设施和技术研究计划，这些中心与与凯克实验室合作，由NHLBI，NIDA，NIAID和NCRR资助。如果这笔赠款得到资助，DIGE系统将是耶鲁社区独有的，它肯定会对生物医学研究做出独特的贡献，远远超出耶鲁。

This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation Grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the grant, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): Funding is requested for a 2-dimensional differential (fluorescence) gel electrophoresis (DIGE) system for the Keck Lab. This leased system is being used very successfully to identify differentially expressed proteins/biomarkers in conjunction with an existing nano-LC-ESI-QTOF for protein identification. DIGE is the most cost effective, powerful, and productive protein profiling technology in the Keck Lab and it is being heavily used to analyze a wide range of experimental versus control samples from whole cells, partially purified protein extracts, and biological fluids like sera. Our DIGE system has been used to carry out >350 quantitative analyses for 48 Yale and 32 non-Yale investigators at 19 institutions. Since entering into a lease in December, 2003, demand for DIGE has continued to increase. Improvements made with the requested, newer version image analysis software and our newly installed, high throughput and sensitive MALDI TOF/TOF mass spectrometer will make this technology state-of-the-art, and these improvements will further increase demand by investigators studying fundamental mechanisms of diseases including autism, substance abuse, cerebral palsy, mental retardation, premature birth, asthma, type I diabetes, renal diseases, tumor cell resistance, genesis of myeloid leukemia, molecular bone development, and Lupus. If this grant is not awarded, the 4-fold increase in monthly lease payments (as the lease matures from interest only to interest + capital equipment cost repayments starting in May, 2006) within the next year will result in a very substantial increase in DIGE service charges. Such an increase would very negatively impact on use of this advanced technology by both the Yale and non-Yale scientific community. Strengths of this application include the experience of the co-Pi overseeing large, multi-investigator NIH contracts and Centers; the complementary biotechnological, statistical, and database expertise of the principal investigator and other key personnel; the extensive protein profiling technologies already in use in the Keck Lab, and the strong infrastructure and technological research programs supported by several proteomics and high performance computing centers that are closely associated with the Keck Lab and are funded by NHLBI, NIDA, NIAID, and NCRR. If this grant is funded, the DIGE system would be unique to the Yale Community, and it surely would make a unique contribution to biomedical research that would extend far beyond Yale.