CORE--PATIENT MUTATION ANALYSIS

核心——患者突变分析

• 批准号: 6242874
• 负责人: EILEEN REMOLD-O'DONNELL
• 金额: $ 38.11万
• 依托单位: IMMUNE DISEASE INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-30 至 1998-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6242874
• 关键词: Wiskott Aldrich syndrome; biomedical facility; blood cells; cell line; cell population study; cryopreservation; gene expression; gene mutation; human tissue; lymphocyte; nucleic acid sequence; phenotype; single strand conformation polymorphism; tissue /cell culture; tissue resource /registry; western blottings

The Patient Mutation Analysis Core, is a blood processing, tissue culture and mutation analysis core. In this facility, we collect and process peripheral blood of WAS and XLT patients including quantifying and phenotyping blood cell subpopulations, isolation of mononuclear cells and lymphocytes, cryopreservation, and generation, maintenance and characterization of EBV-transformed B-cell lines that will be used in investigations in Projects 1 and 2. The Core contributes to diagnosis of WAS in newly referred patients by performing platelet sizing and, where family history is lacking, heterozygosity analysis of female relatives. The Core identifies the WAS gene mutation of diagnosed patients by isolating DNA and amplifying exon regions, analyzing amplified regions by single chain conformation polymorphism analysis (SCCP) followed be sequencing of the abnormal region. The Core characterizes and quantifies the mutated WASP protein expressed in patient cells by a combination of Western blotting and Phosphor-Imaging. The Core will determine whether addition of protease inhibitors, a potential therapeutic treatment, will increase the level of WASP protein in cultured cells of some patients. Finally the Core maintains a database of patient blood samples and cell lines as well as clinical and molecular information on participating patients and a separate database of published WAS defects.

患者突变分析的核心，是血液加工、组织培养 和突变分析核心。在这个设施里，我们收集和加工 心力衰竭和心力衰竭患者外周血的检测 血细胞亚群的表型，单核细胞的分离和 淋巴细胞，冷冻保存，以及产生，维持和 EBV转化的B细胞系的特性将用于 对项目1和项目2的调查。核心有助于诊断 是在新转诊的患者中进行血小板大小测定，其中 缺乏家族史、女性亲属杂合性分析。 Core通过以下方式识别诊断患者的as基因突变 提取DNA并扩增外显子区域，对扩增区域进行分析 单链构象多态分析(SCCP) 对异常区域进行测序。核心描述和量化 突变的WASP蛋白在患者细胞中通过结合 蛋白质印迹和荧光成像。核心将决定是否 添加蛋白酶抑制剂，一种潜在的治疗方法，将 提高部分患者培养细胞中WASP蛋白水平。 最后，核心维护一个病人血液样本和细胞的数据库 LINES以及参与的临床和分子信息 患者和另一个已发表的数据库存在缺陷。