CORE--CELL /MOLECULAR /MICROSCOPY

核心--细胞/分子/显微镜

• 批准号: 6951265
• 负责人: THOMAS TOMASELLI
• 金额: $ 26.87万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-15 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6951265
• 关键词: bioinformatics; biomedical facility; cardiovascular disorder therapy; cell biology; electronic pacemaker; gel electrophoresis; gene expression; heart conduction system; heart contraction; heart disorder; heart electrical activity; heart failure; immunocytochemistry; liquid chromatography; mass spectrometry; microarray technology; microscopy; molecular biology; molecular biology information system; molecular genetics; nucleic acid purification; posttranslational modifications; protein quantitation /detection; proteomics

The molecular genetics and proteomics core serves as a resource to provide expertise and assistance in the analysis of gene and protein expression to projects by Van Eyk and Tomaselli. The emphasis of the core will be on the preparation and utilization of mRNA and protein for the performance and analysis of microarray and protein expression studies. Activities of the core facility will be performed in several locations on the campus of the Johns Hopkins School of Medicine including laboratories in the Division of Cardiology, the Sidney Kimmel Comprehensive Cancer Care Microarray Facility and the Center for Computational Medicine and Biology centered in the Department of Biomedical Engineering. The services provided by this Core include: mRNA preparation and qualification for kinetic real time PCR (RT-PCR) and microarray experiments; access to canine EST libraries; spotting and hybridization to microarrays, assistance with analysis of microarray data; access to CAGE, a database repository for microarray experiments; primers for RT-PCR; canine tissue qualified antibodies and protocols for their use in Western blotting, immunohistochemical and immunocytochemical applications; cytosolic, sarcolemmal, sarcoplasmic reticulum, ERK1/2 and connexin subproteome preparation and analysis (including the performance of 2D gel electrophoresis (2DE), 2D liquid chromatography (2DLC), differential imaging gel electrophoresis (DICE) and mass spectrometry); and a canine heart-specific protein database for peptide mass fingerprinting (PMF). In addition to the provision of technical expertise to the projects, the core will generate significant cost savings by serving as a centralized resource for molecular biological and protein chemical analysis of the canine heart.

分子遗传学和蛋白质组学核心作为一个资源，提供专业知识， 协助货车Eyk和Tomaselli的基因和蛋白质表达分析项目。核心的重点将是对mRNA和蛋白质的制备和利用，用于微阵列和蛋白质表达研究的性能和分析。核心设施的活动将在约翰霍普金斯医学院校园内的几个地点进行，包括心脏病科的实验室、Sidney Kimmel综合癌症护理微阵列设施和以生物医学工程系为中心的计算医学和生物学中心。该核心提供的服务包括：mRNA的制备和动态真实的时间PCR（RT-PCR）和微阵列实验的资格;进入犬EST库;斑点和杂交到微阵列，协助分析微阵列数据;进入CAGE，微阵列实验的数据库; RT-PCR引物;犬组织合格抗体及其在Western印迹、免疫组织化学和免疫细胞化学应用中的使用方案;细胞质、肌膜、肌浆网、ERK 1/2和连接蛋白亚蛋白质组制备和分析（包括2D凝胶电泳（2DE）、2D液相色谱（2DLC）、差示成像凝胶电泳（DICE）和质谱的性能）;和用于肽质量指纹（PMF）的犬心脏特异性蛋白质数据库。除了为项目提供技术专门知识外，核心还将作为犬心脏分子生物学和蛋白质化学分析的集中资源，节省大量费用。