CONSTRUCTION OF A TARGETED RHESUS MACAQUE MICROARRAY

描述(由申请人提供)： 恒河猴(Macaca Mulatta)在解剖学和生理学上与人类相似，为生物医学研究提供了独特的机会。针对艾滋病毒和可能的生物恐怖分子，如天花和炭疽的疫苗开发需要使用非人类灵长类动物(NHP)。恒河猴也是研究生殖生物学和高级皮质功能的首选动物。这些领域的重要研究目前因缺乏关键工具--表达微阵列而受阻。这项提议的目标是利用人类基因组计划的信息，通过对人类基因的猴子同源物进行定向聚合酶链式反应(PCR)扩增来创建恒河猴表达微阵列。为了最大限度地发挥科学和生物医学的影响，这一有针对性的方法将分三个有序的阶段优先考虑。第一阶段将产生一个由1800个基因组成的有重点的阵列，NHP社区将其选为最优先考虑的基因。第二个阶段将是识别和放大那些人类探针效果不佳的猴子基因。第三个阶段是扩增剩下的恒河猴基因。已经从恒河猴cdna文库克隆的基因将被排除在这个项目之外。每个阶段将有四个步骤：(1)鉴定基因；(2)设计用于定向扩增这些基因的引物对；(3)扩增、亚克隆、测序、注释和储存这些基因克隆；以及(4)构建靶向恒河猴表达微阵列。该项目将产生一种对国家热带雨林社区非常重要的资源。

DESCRIPTION (provided by applicant): The rhesus macaque (Macaca mulatta) offers unique opportunities for biomedical research due to similarities to humans in both anatomy and physiology. Vaccine development for HIV and possible bioterrorist agents such as small pox and anthrax require the use of nonhuman primates (NHPs). The rhesus macaque is also the animal of choice for studies of reproductive biology and higher cortical functions. Important studies in these areas are currently impeded by the lack of a critical tool - expression microarrays. The objective of this proposal is to create rhesus macaque expression microarrays by targeted polymerase chain reaction (PCR) amplification of monkey homologs of human genes using information from the human genome project. To maximize scientific and biomedical impact, this targeted approach will be prioritized in three ordered stages. The first stage will be to produce a focused array of 1,800 genes chosen as highest priority by the NHP community. The second stage will be to identify and amplify those monkey genes for which human probes work poorly. The third stage will be to amplify the remaining rhesus macaque genes. Genes already cloned from rhesus cDNA libraries will be excluded from this project. Each stage will have four steps: (1) identify the genes; (2) design primer pairs for targeted amplification of those genes; (3) amplify, subclone, sequence, annotate, and deposit those gene clones; and (4) construct targeted rhesus expression microarrays. This project will produce a resource of major importance to the NHP community.