411POWRE: Analysis of Microbial Communities by Coupled Polymerase Chain Reaction and Dematuring Gradient Gel Electrophoresis as an Indicator of in Situ Bioremediatio

9870561 Marr本研究的目的是为原位生物修复的验证奠定基础。该研究将验证聚合酶链反应（PCR）和变性梯度凝胶电泳（DGGE）分子生物学工具可用于监测与氯化溶剂原位生物降解相关的微生物种群演替的假设。PCR和DGGE已成功应用于天然微生物群落，以阐明微生物多样性，并确定未受污染的水生、沉积物和土壤环境中存在的特定种群。具体目标是：1)优化PCR和DGGE的应用，以区分未污染的甲烷营养化土壤中的微生物种类；2)同时监测三氯乙烯降解和物种多样性，通过PCR扩增的16S rDNA的DGGE分辨率测量；3)通过类型特异性系统发育和分解代谢基因探针以及16S rDNA序列分析，随着时间的推移鉴定优势物种。本研究的最终影响将是提高我们对生物修复的认识，并提供工具来监测和优化污染地下水现场生物修复的操作条件。该项目将使PI在消毒过程领域的研究人员中处于领导地位，将她的研究重点放在一个存在重大知识空白的领域

9870561 Marr The objective of this research is to establish a foundation for the verification of in situ bioremediation. The proposed research will test the hypothesis that the molecular biology tools of polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE) can be used to monitor the succession of microbial populations associated with in situ biodegradation of chlorinated solvents. PCR and DGGE have been successfully applied to natural microbial communities to elucidate microbial diversity and identify the specific populations present in uncontaminated aquatic, sediment and soil environments. Specific objectives are to 1) optimize the application of PCR and DGGE to distinguish microbial species in uncontaminated methanotrophic soil, 2) simultaneously monitor trichloroethylene degradation and species diversity as measured by DGGE resolution of PCR-amplified 16S rDNA and 3) identify the predominant species over time via type-specific phylogenetic and catabolic gene probes and by 16S rDNA sequence analysis. The ultimate impact of this research will be to enhance our knowledge of bioremediation and to provide tools to monitor and optimize operating conditions during in situ bioremediation of contaminated groundwater sites. This project will allow the PI to take a leadership position among researchers in the area of disinfection processes by focusing her research in an area were there are significant knowledge gaps