Molecular Assessment of Hydrocarbon Degradation

碳氢化合物降解的分子评估

基本信息

批准号：
6803237
负责人：
GERBEN J ZYLSTRA
金额：
$ 23.33万
依托单位：
RUTGERS, THE STATE UNIV OF N.J.
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-09-30 至 2006-07-31
项目状态：
已结题

项目摘要

The primary goal of this project is to obtain information about community function by focusing on the critical initial oxygenases that are the rate-limiting step in productive degradation and eventual mineralization of aromatic compounds in oxic and suboxic environments. This project will explore the diversity of oxygenase gene sequences and monitor shifts in their population without regard to host organism. Results of the proposed experiments will identify the breadth of genetic diversity of oxygenases in the environment and associate that diversity with known and unknown organisms. In particular the data will identify gene targets for better molecular assessment of hydrocarbon oxidation in the environment. From this two-year exploratory research program we anticipate the development of a large database of oxygenase gene sequences. It is anticipated that the majority of these will be novel sequences, derived from organisms that are not yet in laboratory cultivation. Our ability to successfully obtain such sequences will result from the use of novel oxygenase PCR primers. Full length oxygenase gene sequences will be obtained through inverse PCR and sequencing of environmental DNA cosmid clones. The goal of developing and using these molecular approaches is to assess their robustness for tracking changes in oxygenase gene populations in laboratory-scale environmental perturbation experiments or in remediation treatment applications. This will be accomplished by focusing intensively on source materials from a single Superfund site in which differences in physical, chemical and geological site parameters can be assessed for their impact on the "molecular fingerprints" of the microbial oxygenase patterns that result from the experimental treatments. Data from this type of exploratory study should allow us to determine which molecular signatures (in this case, dioxygenase patterns) are predictive of overall biodegradative perforrnance of the system. From this, we plan to construct a new generation of "oxygenase microarray" that can provide functional information about those members of the microbial community that become important in biodegradative and bioremediative processes.

该项目的主要目标是通过关注关键的初始加氧酶来获取有关群落功能的信息，这些初始加氧酶是含氧和缺氧环境中芳香族化合物生产性降解和最终矿化的限速步骤。该项目将探索加氧酶基因序列的多样性，并监测其种群的变化，而不考虑宿主生物体。拟议实验的结果将确定环境中加氧酶遗传多样性的广度，并将这种多样性与已知和未知的生物体联系起来。特别是，这些数据将确定基因目标，以便更好地对环境中的碳氢化合物氧化进行分子评估。通过这个为期两年的探索性研究计划，我们预计将开发出一个大型加氧酶基因序列数据库。预计其中大部分将是新序列，源自尚未在实验室培养的生物体。我们成功获得此类序列的能力将归功于新型加氧酶 PCR 引物的使用。通过环境DNA粘粒克隆的反向PCR和测序可以获得全长加氧酶基因序列。开发和使用这些分子方法的目标是评估它们在实验室规模的环境扰动实验或修复处理应用中跟踪加氧酶基因群变化的稳健性。这将通过集中关注来自单个超级基金站点的源材料来实现，其中可以评估物理、化学和地质站点参数的差异对实验处理产生的微生物加氧酶模式的“分子指纹”的影响。来自此类探索性研究的数据应该使我们能够确定哪些分子特征（在本例中为双加氧酶模式）可以预测整体生物降解系统的性能。由此，我们计划构建新一代“加氧酶微阵列”，它可以提供有关在生物降解和生物修复过程中变得重要的微生物群落成员的功能信息。