REGULATION OF EXPRESSION OF CLASS I MHC GENE

I 类 MHC 基因表达的调节

• 批准号: 3916415
• 负责人: D SINGER
• 金额: --
• 依托单位: DIVISION OF CANCER BIOLOGY AND DIAGNOSIS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3916415
• 关键词: chemical binding; chromosome deletion; gene expression; genetic mapping; genetically modified animals; histocompatibility antigens; immune response genes; laboratory mouse; miniature swine; regulatory gene

The aim of this work is to investigate the mechanisms controlling the expression of a multi-gene family. The miniature swine has been chosen as an experimental model because there are only 7 members of the family. To address the question of the molecular regulation of the expression of class I MHC genes, two approaches have been taken: 1) the analysis of in vivo patterns of expression of each of the genes, both in situ and in transgenic mice; and 2) characterization of regulatory elements associated with these genes. Analysis of in vivo patterns reveals that both shared and discrete regulatory mechanisms operate within the family. Thus, some genes are coordinately expressed, whereas others are not. In both cases, patterns of expression are actively regulated. Regulatory sequences within one of the transplantation antigen genes have been identified by generating a series of 5' end deletion mutants. A series of both positive and negative elements have been mapped. A novel negative regulatory element, which requires the presence of a positive element in order to function, has been identified. It has been further demonstrated that these elements function through the binding of tran sacting factors. The binding sites of the trans acting factors have been mapped using both gel retardation assays and exonuclease protection.

这项工作的目的是研究控制机制 多基因家族的表达。 迷你猪 被选为实验模型，因为只有7个 家庭成员。 为了解决分子问题 调节I类MHC基因的表达，两种方法 已采取：1）在体内表达模式的分析 在原位和转基因小鼠中的每种基因;以及2） 与这些相关的调控元件的表征 基因. 体内模式的分析表明，共享和离散 家庭内部的管理机制发挥作用。 因此，一些基因 是协调表达的，而另一些则不是。 在这两种情况下， 表达模式受到积极调控。 移植抗原之一内的调节序列 已通过产生一系列5'末端来识别基因 缺失突变体 一系列积极和消极的因素 已经被映射了。 一种新的负调控元件， 需要一个积极的因素来发挥作用， 已经被确认了 进一步证明，这些 元素通过结合转化因子发挥作用。 的 反式作用因子的结合位点已经用 凝胶阻滞测定和核酸外切酶保护。