Genomics of High Temperature Biological Systems

描述（由申请人提供）：第三界的许多成员，古细菌，是极端的超嗜热生物。这些生物在分子生物学中提出了许多有趣的基本问题，因为它们所有的生命过程都发生在高温下。例如，在100摄氏度以上的温度下，调节蛋白是如何识别和结合操作员的？DNA如何以高保真度复制，这些生物体如何避免由于热诱导的DNA损伤而过度突变，以及它们采用什么修复策略？我们建议对高温古菌之一的嗜气焦杆菌（Pyrobaculum aerophilum）进行功能基因组分析，该细菌可以生长到104℃，最佳生长温度为100℃。我们已经对嗜气焦杆菌（Pyrobaculum aerophilum）的2.2百万碱基基因组进行了测序和完全注释，并正在与斯坦福大学/加州大学加州大学的Todd Lowe合作完成包含每个ORF的微阵列的构建圣克鲁斯。我们将开展不同环境下全基因组基因表达实验，利用数据鉴定共调控基因。通过扩展分析，我们希望利用数据建立实验，以确定调控途径和调控蛋白。我们将扩展遗传系统的发展，目的是能够在这种有机体中进行反向遗传。我们已经开始深入研究这种生物的修复系统，我们将继续研究DNA修复策略和蛋白质。

DESCRIPTION (provided by applicant): Numerous members of the third kingdom, Archaea, are extreme hyperthermophiles. Such organisms pose many interesting fundamental questions in molecular biology, since all of their life processes occur at elevated temperatures. For instance, how do regulatory proteins recognize and bind to operators at temperatures above 100 degrees C? How can the DNA replicate with high fidelity, and how do these organisms avoid excessive mutation due to heat-induced DNA lesions, and what repair strategies do they employ? We propose to develop a functional genomic analysis of one of the high temperature archaea, Pyrobaculum aerophilum, that can grow up to 104 degrees C, with an optimal growth temperature of 100 degrees C. We have sequenced and fully annotated the 2.2 mega base genome of Pyrobaculum aerophilum, and are completing the construction of microarrays containing every ORF, in collaboration with Todd Lowe at Stanford/U.C. Santa Cruz. We will carry out experiments on genome-wide gene expression in different environments, and use the data to identify coregulated genes. Through extended analyses, we hope to use the data to set up experiments to identify regulatory pathways and regulatory proteins. We will extend the development of a genetic system with the aim of being able to carry out reverse genetics in this organism. We have begun an indepth investigation of the repair systems in this organism, and we will continue to characterize DNA repair strategies and proteins.