Functional Genemics of Cryptosporidium parvum

描述（由申请人提供）：这是一份修订后的资助申请，用于研究小隐孢子虫和感染该寄生虫的宿主细胞中的全局基因表达。小隐孢子虫是艾滋病患者的机会性病原体，是全世界儿童腹泻的常见原因。细小疟原虫基因组的完整序列为确定药物靶点以及在基因组和功能水平上对该寄生虫的研究提供了新的机会。在这个项目中，DNA微阵列将用于研究寄生虫基因调控和宿主细胞对这种感染的反应。分析寄生虫的全局转录，主要是在脱胞、宿主细胞入侵和细胞内发育的初始阶段，将确定寄生虫的生化途径，这些途径在生命周期的这一阶段有差异表达。在培养和动物模型中分析受感染宿主细胞的转录反应将确定宿主对感染的反应机制，并阐明导致隐孢子虫病肠道症状的分子途径。该项目是两个研究小组之间的合作努力，他们在小孢子虫的生物学和基因组学方面具有互补的专业知识和广泛的合作经验。

DESCRIPTION (provided by applicant): This is a revised grant application to study global gene expression in Cryptosporidium parvum and in host cells infected with this parasite. Cryptosporidium parvum is an opportunistic pathogen in people with AIDS and is a common cause of diarrhea in children worldwide. The completed sequence of the C. parvum genome provides new opportunities for identifying drug targets and for research on this parasite on a genomic and functional level. In this project DNA microarrays will be used to study parasite gene regulation and the response of the host cell to this infection. Analysis of global parasite transcription, primarily during excystation, host cell invasion, and the initial phase of intracellular development will identify parasite biochemical pathways, which are differentially expressed during this phase of the life cycle. Analysis of the transcriptional response of infected host cells in culture and in animal models will identify mechanisms by which the host responds to the infection, and shed light on the molecular pathways leading to the intestinal symptoms of cryptosporidosis. This project is a collaborative effort between two research groups with complementary expertise in the biology and genomics of C. parvum and extensive collaborative experience. In Specific Aim 1 a comprehensive DNA microarray containing all of the genes of C. parvum will be developed. This task will take advantage of the genomic sequence by converting genomic information into a tool for identifying genetic processes associated with parasite development. In Specific Aim 2 microarrays developed under aim 1 will be used to probe global gene expression during oocyst excystation, host cell invasion, and first generation merogony. Specific Aim 3 will use commercially available human and mouse gene arrays to study the response of the host cell to C. parvum. A comparison of the host cell response to C. parvum type 1 and type 2 will be performed to probe the molecular basis of the observed difference in virulence between types.