DIFFERENTIAL GEL ELECTROPHORESIS (DIGE) SYSTEM: AUTISM

这个子项目是利用由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。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是用于拨款的，而不一定是用于调查人员的机构。描述(由申请人提供)：为凯克实验室的二维差示(荧光)凝胶电泳(DGE)系统申请资金。这个租赁的系统正在非常成功地与现有的用于蛋白质鉴定的纳米LC-ESI-QTOF一起用于鉴定差异表达的蛋白质/生物标记物。DGE是Keck实验室中最具成本效益、功能最强大和最具生产力的蛋白质图谱技术，它正被广泛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的各种实验样品和对照样品。我们的DGE系统已被用于对19个机构的48名耶鲁调查人员和32名非耶鲁调查人员进行&gt；350定量分析。自2003年12月签订租约以来，对DIGE的需求持续增加。根据要求，更新版本的图像分析软件和我们新安装的高通量、高灵敏度的MALDI TOF/TOF质谱仪所做的改进将使这项技术成为最先进的技术，这些改进将进一步增加研究人员对研究疾病基本机制的需求，这些疾病包括自闭症、药物滥用、脑瘫、智力低下、早产、哮喘、I型糖尿病、肾脏疾病、肿瘤细胞耐药、髓系白血病的发生、分子骨发育和狼疮。如果不授予这笔赠款，下一年内每月租赁付款(由于租赁从2006年5月开始从仅计利息到期到利息+资本设备成本偿还)增加4倍，将导致DGE服务费大幅增加。这样的增长将对耶鲁和非耶鲁科学界使用这项先进技术产生非常负面的影响。这一应用的优势包括共同PI监督大型、多研究员NIH合同和中心的经验；首席研究员和其他关键人员的互补性生物技术、统计和数据库专业知识；Keck实验室已经使用的广泛的蛋白质图谱技术，以及由几个与Keck实验室密切相关并由NHLBI、NIDA、NIAID和NCRR资助的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。如果这笔资金得到资助，DGE系统将是耶鲁社区独一无二的，它肯定会为生物医学研究做出独特的贡献，远远超出耶鲁的范围。

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.