Project 4: Application of Comparative Genomics, Transcriptomics, & Proteomics Opt

项目4：比较基因组学、转录组学的应用，

• 批准号: 7089426
• 负责人: LISA ALVAREZ-COHEN
• 金额: $ 38.61万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7089426
• 关键词: anaerobiosis; bacterial genetics; biotransformation; chemical kinetics; chlorination; detoxification; environmental exposure; environmental toxicology; gene expression; genetic regulatory element; genetic transcription; mass spectrometry; microarray technology; polymerase chain reaction; protein biosynthesis; proteomics; trichloroethylene

This proposal will apply advanced molecular tools to understand and optimize the microbial detoxification of common Superfund pollutants, perchloroethene (PCE) and trichloroethene (TCE), which pose a significant threat to human and ecological health. By studying the fundamental processes responsible for anaerobic microbial degradation of PCE and TCE, this work will promote improved situ bioremediation processes. Previous NIEHS-funded research sought to identify and understand the microbes within complex communities responsible for dechlorination. This work takes the next logical step by focusing on the only genus of bacteria, Dehalococcoides, known to completely reduce PCE and TCE to ethene. Whole-genome microarrays, proteomic analyses, and quantitative PCR will be used to characterize the genomic differences between a variety of Dehalococcoides strains and to evaluate gene expression and proteomic changes caused by reductive dechlorination of a variety of substrates, growth in simple and complex microbial communities, and other physiological perturbations. Genomic and transcriptomic comparison of Dehalococcoides strains with different degradation abilities will identify the pathways responsible for specific and general metabolism as well as reveal the evolutionary relationship between the various isolated strains. Transcriptomic comparison of Dehalococcoides strains in pure and mixed cultures will identify pathways involved in inter-species interactions, reveal the nutritional needs and metabolic roles of Dehalococcoides in consortia, and address the limitation in bioremediation applications presented by the poor growth of isolated Dehalococcoides strains. Data from strain identification, gene.expression, and protein production will be complied into kinetic models that can be used to predict rates of reductive dechlorination by poorly characterized microbial communities. This research will meet the SBRP goal of limiting the human exposure and toxicity of chemicals commonly found at Superfund sites by advancing the development of in situ bioremediation of PCE and TCE, a technology that destroys contaminants in their subsurface location without extraction to the surface, avoiding potential human and ecological exposure.

该建议将应用高级分子工具来了解和优化微生物 普通超级基金污染物，高氯乙烯（PCE）和三氯乙烯（TCE）的排毒 对人类和生态健康构成重大威胁。通过研究基本过程 负责PCE和TCE的厌氧微生物降解，这项工作将促进原位改善 生物修复过程。以前由NIEHS资助的研究试图识别和理解 复杂社区中的微生物负责脱氯。这项工作采取了下一个逻辑步骤 通过关注唯一的细菌属，去足球，已知可以将PCE和TCE完全降低至 乙烯。全基因组微阵列，蛋白质组学分析和定量PCR将用于表征 多种去足球菌菌株之间的基因组差异并评估基因表达 以及由各种底物的脱氯的氯化引起的蛋白质组学变化，简单和 复杂的微生物群落和其他生理扰动。基因组和转录组 与不同降解能力的去足球菌菌株的比较将识别途径 负责特定和一般的新陈代谢，并揭示 各种孤立的菌株。纯培养和混合培养物中去丘脑球菌菌株的转录组比较 将确定涉及种间相互作用的途径，揭示营养需求和代谢角色 联盟中的去核苷的dehalocoides，并解决了生物修复应用的限制 孤立的脱核苷菌株的生长差。来自菌株识别，基因表达和 蛋白质产生将被遵守动力学模型，可用于预测还原的速率 微生物群落特征不佳的氯化。这项研究将达到SBRP的目标 通过推进，限制在超级基金站点常见的化学物质的接触和毒性 PCE和TCE的原位生物修复的开发，这项技术会破坏其污染物 地下位置没有提取表面，避免了潜在的人类和生态暴露。