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

9870561 MARR这项研究的目的是为现场生物修复的验证奠定基础。这项拟议的研究将检验这样一种假设，即聚合酶链式反应(PCR)和变性梯度凝胶电泳(DGGE)这两种分子生物学工具可以用来监测与氯化溶剂原位生物降解相关的微生物种群的演替。PCR和DGGE已被成功地应用于自然微生物群落，以阐明微生物多样性，并识别存在于未受污染的水、沉积物和土壤环境中的特定种群。具体目标是1)优化应用PCR和DGGE来区分未受污染的甲烷营养土壤中的微生物物种，2)同时监测三氯乙烯的降解和物种多样性，通过PCR扩增的16S rDNA的DGGE分辨率来测量，以及3)通过特定类型的系统发育和分解代谢基因探针和16S rDNA序列分析来识别随着时间的推移优势物种。这项研究的最终影响将是增强我们对生物修复的知识，并提供工具来监测和优化受污染地下水现场生物修复过程中的运行条件。该项目将使PI在消毒过程领域的研究人员中处于领先地位，将她的研究集中在一个存在重大知识差距的领域

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