TRANSCRIPTION OF ACANTHAMOEBA RNA POLYMERASE II GENES

基因表达的调控是细菌生长发育的核心。 所有的有机体。扰乱正常的控制机制会导致细胞 死亡或蜕变。在其他情况下，颠覆细胞 转录装置可以构成病毒感染的基础。 适当地，要了解真核转录的机制是 现代生物学的一个主要目标。 这是一项研究沙门氏菌rna聚合酶II基因转录的建议。 模式生物卡氏棘阿米巴。长期目标是 在分子水平上建立真核基因表达的原理。 卡氏棘阿米巴肌动蛋白基因的体内结构 要下定决心。结果将确定相互作用的启动子区域 并将作为体外研究的参照点 学习。 棘阿米巴体外转录系统的研制 使用棘阿米巴肌动蛋白I和肌球蛋白II基因的卡氏杆菌。推动者 肌动蛋白基因体外准确转录所需的元件 将被确定，以及所需因素的数量和类型 笔录将会确定。因子之间的相互作用，RNA 聚合酶II和DNA将通过足迹法进行检查。步骤如下： 中定义了导致引发的预引发复合体组件 聚合酶和因子结合以及DNA解离或其他术语 构象变化。转录因子的纯化将是 使用常规亲和力和启动子亲和力的组合启动 方法：研究方法。

The control of gene expression is central to the growth and development of all organisms. Disrupting the normal control mechanisms can lead to cell death or transformation. In other cases, subverting the cells transcription apparatus can form the basis for viral infection. Appropriately, to understand the mechanisms of eukaryotic transcription is a major goal in modern biology. This is a proposal to study transcription of RNA polymerase II genes from the model organism Acanthamoeba castellanii. The long-term goal is to establish principles of eukaryotic gene expression at the molecular level. The in vivo structure of an actin gene from Acanthamoeba castellanii will be determined. The results will identify promoter regions that interact with protein in vivo and will be used as a reference point for in vitro studies. An in vitro transcription system will be developed form Acanthamoeba castellanii using the Acanthamoeba actin I and myosin II genes. Promoter elements required for accurate transcription in vitro from the actin gene will be identified and the number and type of factors required for transcription will be determined. The interaction between factors, RNA polymerase II and DNA will be examined by footprinting. The steps of preinitiation complex assembly that lead to initiation will be defined in terms of polymerase and factor binding as well as DNA unwinding or other conformational changes. Purification of transcription factors will be initiated using a combination of conventional and promoter affinity methods.