Molecular Assessment of Hydrocarbon Degradation

碳氢化合物降解的分子评估

基本信息

批准号：
6743011
负责人：
GERBEN J ZYLSTRA
金额：
$ 19.44万
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 2005-07-31
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.

该项目的主要目的是通过关注关键的初始氧化酶来获取有关社区功能的信息，这些酶是氧化和亚氧化环境中芳族化合物的生产降解和最终矿化的限制步骤。该项目将探索氧合酶基因序列的多样性并监测其人群中的变化，而无需考虑宿主生物体。提出的实验的结果将确定环境中氧酶的遗传多样性的广度，并将多样性与已知和未知生物相关联。特别是，数据将确定基因靶标，以更好地评估环境中烃氧化的分子评估。从这个为期两年的探索性研究计划中，我们可以预期开发大型氧合酶基因序列的数据库。可以预料，其中大多数将是新的序列，这些序列源自尚未实验室种植的生物。我们成功获得此类序列的能力将由新型的氧合酶PCR引物产生。全长氧合酶基因序列将通过逆PCR和环境DNA宇宙克隆的测序获得。开发和使用这些分子方法的目的是评估其在实验室规模的环境扰动实验中或修复治疗应用中跟踪氧合酶基因种群变化的鲁棒性。这将通过强烈关注单个超级基金位点的源材料来实现，在这种材料中，可以评估物理，化学和地质位点参数的差异，以评估它们对由实验处理导致的微生物氧酶模式的“分子指纹”的影响。此类探索性研究的数据应使我们能够确定哪些分子特征（在这种情况下为二加氧酶模式）可预测总体生物降解系统的完美。由此，我们计划构建新一代的“氧合酶微阵列”，该新一代可以提供有关微生物社区成员在生物降解和生物补充过程中变得重要的功能信息。