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.

该项目的目的是开发和利用寡核苷酸微阵列技术用于模型丝状真菌粗糙脉孢菌。这些研究将为丝状真菌的基因表达谱提供坚实的基础，从而促进该类群内以及真菌与其他生物之间的比较研究。可用于脉孢菌的丰富的生物和分子工具基础得到了存档质量的基因组序列的补充，从而为在全球范围内提出基因组调控问题创造了机会。 本提案中描述的通过微阵列技术生成的基因表达数据可以显着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代表粗糙链孢霉每个预测基因的 70-met 寡核苷酸的载玻片，并使用这些载玻片标准化我们的技术，以开发微阵列数据的可靠统计分析。我们的第二个目标是在暴露于各种生长条件和环境压力后，对已测序的脉孢菌菌株进行基线转录分析。我们的第三个目标将使用本提案的项目 #1 生成的敲除菌株来测试源自微阵列实验的假设。预期的敲除将包括 (1) 已在粗糙猪笼草中注释的基因，这些基因是 参与酿酒酵母转录调控过程并已进行转录分析的基因，(2) 被注释为酿酒酵母中不存在的推定调控因子的基因，(3) 酿酒酵母和酿酒酵母中不存在的基因的同源物. 粟酒裂殖酵母和在粗糙脉孢菌中具有未知的生物学功能，以及（4）明显的丝状真菌特异性基因，在其他生物体中没有同源物。 该项目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之外，这些数据还将加快对包括重要人类病原体在内的一般真菌的功能分析。