MOLECULAR ANALYSIS OF THE WERNERS GENE PRODUCT

维尔纳基因产物的分子分析

• 批准号: 2769278
• 负责人: ROBERT Anthony MARCINIAK
• 金额: $ 10.31万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2769278
• 关键词: Werner's syndrome; affinity chromatography; aging; confocal scanning microscopy; cytogenetics; gene expression; gene mutation; human genetic material tag; human tissue; image enhancement; immunocytochemistry; immunoprecipitation; laboratory rabbit; loss of heterozygosity; molecular cloning; nucleic acid sequence; nucleotides; phenotype; polymerase chain reaction; protein structure function; single strand conformation polymorphism; tumor suppressor genes; western blottings; yeast two hybrid system

As the demographics Of the United States populace continues to skew towards older age groups, determining the processes that underlie the physical deterioration that occurs in normal aging becomes increasingly relevant. At this time a genetic approach to the analysis of changes that occur with aging is possible and promises to yield new insight into these processes. Recently genes that have been implicated as the causes of disorders of human premature aging have been identified and cloned. Werner's syndrome is one such disorder. The overall aim of this proposal is to use the molecular analysis of the Werner's syndrome protein to better understand the aging process in the normal human population. This analysis will proceed along three interrelated lines of investigation. First, the hypothesis that the specificity of the Werner's protein function is determined by protein-protein interaction will be tested using the method of yeast two-hybrid cloning. Second, the Werner's gene will be expressed in E. coli and polyclonal antiserum produced for an immunocytochemical analysis. Third, novel -Werner's syndrome gene mutations will be identified in spontaneously occurring human sarcomas by analysis for loss of heterozygosity at the Werner's locus. For those tumors showing LOH at the Werner's locus, the presence of mutations in the remaining Werner's sequences will be determined. The candidate is an M.D. and Ph.D., whose medical training was at Harvard Medical School and Ph.D. thesis work was performed with Phillip Sharp at the Massachusetts Institute of Technology. At the time of this award, he will have completed training in internal medicine and subspecialty training in hematology and medical oncology. Although he has had much experience in biochemistry and molecular biology, he has dedicated himself exclusively to rigorous clinical training for the last five years. The work proposed in this application is a considerable departure from his prior experiences, and entering the field of research on the molecular genetics of aging is a substantial career change. The balanced program of mentored research, didactic and career development activities proposed in this application will provide the foundation for a successful career as an independent investigator in the field of the biology of aging.

随着美国人口统计数据的持续倾斜 对老年群体，确定的过程， 在正常衰老中发生的身体退化变得越来越多， 相关的这时一种遗传学的方法来分析变化 是可能的，并有望产生新的见解， 这些过程。最近，一些基因被认为是 人类过早衰老的原因已经确定 克隆。沃纳综合征就是这样一种疾病。总体目标 这项建议的一个重要方面是利用沃纳的分子分析， 综合征蛋白质，以更好地了解衰老过程中的正常 人口。本分析将沿着三个相互关联的 调查路线。首先，假设的特异性， 沃纳蛋白质的功能是由蛋白质-蛋白质相互作用决定的 将采用酵母双杂交克隆的方法进行检测。第二、 Werner's基因将在E.大肠杆菌和多克隆 免疫细胞化学分析用抗血清。三、新颖 - 沃纳综合征基因突变将在自发性 发生人类肉瘤通过分析杂合性丢失在 沃纳的轨迹。对于那些在Werner位点显示洛缺失的肿瘤， 在剩下的沃纳序列中出现突变 测定 候选人是医学博士。和博士学位，他们的医疗 在哈佛医学院接受培训，论文工作是 与菲利普夏普在马萨诸塞州研究所进行的 技术.在这个奖项的时候，他将完成培训， 内科和血液学专科培训， 肿瘤内科虽然他有很多经验， 生物化学和分子生物学，他致力于 严格的临床训练工作 在本申请中提出的是一个相当大的偏离他先前的 经验，并进入分子研究领域， 衰老的遗传学是一个重大的职业变化。平衡方案 指导研究、教学和职业发展活动 本申请中提出的方法将为 作为一名独立调查员， 衰老生物学