Filamentous Fungi Genetic Analysis

丝状真菌遗传分析

• 批准号: 6958181
• 负责人: JENNIFER J. LOROS
• 金额: $ 21.62万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6958181
• 关键词: Neurospora; circadian rhythms; complementary DNA; developmental genetics; functional /structural genomics; fungal genetics; fungal proteins; genetic mapping; genetic polymorphism; genetic transcription; microorganism resource /registry; nucleic acid sequence

The aim of this Project is to develop and utilize oligonucleotide microarray technology for the model filamentous fungus Neurospora crassa. These studies will provide a solid foundation of gene expression profiles for the filamentous fungi that will promote comparative studies within this group and between the fungi and other organisms. The rich foundation of biological and molecular tools available for Neurospora has been complemented with an archival quality genomic sequence, thus creating the opportunity for asking questions of genome regulation on a global scale. Gene expression data generated by microarray technology as described in this proposal can measurably enhance the functional analysis of this organism's genes. Our first objective is to generate slides containing 70-met oligonucleotides representing each of the predicted genes of N. crassa and to use these slides to standardize our techniques to develop sound statistical analysis of microarray data. Our second objective is to carry out base-line transcriptional profiling on the sequenced strain of Neurospora following exposure to a variety of growth conditions and environmental stresses. Our third objective will test hypotheses derived from microarray experiments using knockout strains generated from Project #1 of this proposal. Anticipated knockouts will include (1) genes that have been annotated in N. crassa that are involved in transcriptional regulatory processes in S. cerevisiae and that have been transcriptionally profiled, (2) genes that are annotated as putative regulatory factors that are absent from S. cerevisiae, (3) homologs of genes that are absent from S. cerevisiae and S. pombe and are of unknown biological function in N. crassa, and (4) apparent filamentous fungal-specific genes that do not have homologs in other organisms. The work outlined in this project will provide procedures, computational tools, and baseline transcriptional profiling data for wild type Neurospora grown under a variety of conditions and will pave the way for collaborative studies in the future. In addition to fostering similar studies in other filamentous fungi, these data will expedite the functional analysis of fungi in general including important human pathogens.

本项目的目的是开发和利用模式丝状真菌粗糙脉孢菌的寡核苷酸芯片技术。这些研究将为丝状真菌的基因表达谱提供坚实的基础，这将促进这一群体内以及真菌与其他生物体之间的比较研究。可用于脉孢菌的生物和分子工具的丰富基础已经与档案质量的基因组序列相补充，从而为在全球范围内提出基因组调控问题创造了机会。 如本提案中所述，通过微阵列技术产生的基因表达数据可以显著增强该生物体基因的功能分析。我们的第一个目标是生成包含70-met寡核苷酸的载玻片，这些寡核苷酸代表N. crassa，并使用这些幻灯片来标准化我们的技术，以发展健全的微阵列数据的统计分析。我们的第二个目标是进行基线的链孢霉的测序株暴露于各种生长条件和环境压力下的转录谱。我们的第三个目标是使用本提案项目#1产生的敲除菌株来测试来自微阵列实验的假设。预期的敲除将包括（1）在N. crassa是 参与S.酿酒酵母和已经转录谱的基因，（2）注释为推定的调控因子，不存在于S.（3）酿酒酵母中缺失基因的同源物。酿酒酵母和酿酒酵母。pombe，其生物学功能尚不清楚。crassa，以及（4）在其他生物中没有同源物的明显丝状真菌特异性基因。 该项目中概述的工作将为在各种条件下生长的野生型脉孢菌提供程序，计算工具和基线转录谱数据，并为未来的合作研究铺平道路。除了促进其他丝状真菌的类似研究外，这些数据还将加快真菌的功能分析，包括重要的人类病原体。