MOLECULAR ANALYSIS OF THE WERNERS GENE PRODUCT

随着美国人口结构的持续倾斜 对于年龄较大的群体，确定基础的过程 在正常衰老过程中发生的身体退化变得越来越严重 切合实际。在这个时候用遗传学的方法来分析变化 随着年龄的增长而发生的事情是可能的，并有望为 这些过程。最近，一些基因被认为是 人类早衰疾病的原因已被确定 并进行了克隆。沃纳综合征就是这样一种障碍。总体目标 这一建议的核心是使用沃纳的分子分析 证候蛋白更好地了解正常人群的衰老过程 人类人口。这一分析将沿着三个相互关联的方向进行 调查的路线。首先，假设生物多样性的特殊性 沃纳蛋白质的功能是由蛋白质之间的相互作用决定的 将采用酵母双杂交克隆的方法进行检测。第二, Werner‘s基因将在大肠杆菌和多克隆中表达 为免疫细胞化学分析而产生的抗血清。第三，小说 -Werner综合征基因突变将自发地在 人类肉瘤发生的基因杂合性缺失分析 沃纳轨迹。对于那些在Werner‘s轨迹显示杂合性缺失的肿瘤， 其余Werner序列中突变的存在将是 下定决心。候选人是医学博士和博士，他的体检 培训在哈佛医学院进行，博士论文工作是 与菲利普·夏普在麻省理工学院演出 技术。在获得这一奖项时，他已经完成了训练 在血液学和血液学的内科和专科培训中 内科肿瘤学。尽管他有丰富的经验 生物化学和分子生物学，他致力于 在过去的五年里接受了严格的临床培训。这项工作 在这份申请书中提出的是与他之前的 经验，并进入分子研究领域 衰老的遗传学是一种实质性的职业变化。平衡计划 指导研究、教学和职业发展活动 在本应用程序中提出的将为 作为一名成功的独立调查员，在 衰老生物学。

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.