RENIN EXPRESSING CELL LINES FOR HYPERTENSION RESEARCH

描述：这是一份修订后的申请，旨在调查 肾素基因启动子在肾素表达细胞系As4.1中的表达。这个 申请者将尝试在5‘年内完成顺式代理站点的划定 小鼠REN-1C基因的FS似乎是驱动高密度脂蛋白的关键 报告构建物的水平细胞特异性表达。这将是 利用瞬时转染法在肾素表达中的应用 AS4.1细胞，以及不表达肾素的Ltk细胞。以确保 已确定的元素与体内调控密切相关，关键元素将 在转基因小鼠身上进行功能测试。常规化验方法 DNA-蛋白质的相互作用将被用来细化个体的极限 监管地点，并将作为识别的基础， 结合蛋白的转录调控因子的纯化和克隆 这些网站。这种蛋白质一旦拿到手，将被表征为 相关功能结构域及其发育表达模式将 在正常和病理生理条件下进行检查以确定 这些模式是否兼容，从时间顺序和 地形学观点，与肾素基因转录调控。

DESCRIPTION: This is a revised application to investigate the regulation of the renin gene promoter in a renin-expressing cell line, As4.1. The applicant will attempt to complete delineation of cis-acting sites in the 5' FS of the murine Ren-1c gene that appear to be critical for driving high level cell-specific expression of reporter constructs. This will be accomplished utilizing transient transfection assays in the renin-expressing AS4.1 cells, as well as the non-renin-expressing Ltk- cells. To ensure that the elements identified are germane to regulation in vivo, key elements will be tested for function in transgenic mice. Conventional assays of DNA-protein interactions will be used to refine the limits of the individual regulatory sites and will serve as a basis for the identification, purification and cloning of regulatory transcription factors which bind to these sites. Such proteins, once in hand, will be characterized for relevant functional domains and their developmental expression patterns will be examined under normal and pathophysiological conditions to determine whether these patterns are compatible, from a chronological and topographical standpoint, with regulation of renin gene transcription.