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| 实时荧光PCR在治理石油污染中的应用 |
| Application of Real Time PCR in Harnessing Oil Pollution |
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| DOI：doi:10.3969/j.issn.1005-7021.2014.02.007 |
| 中文关键词: 枯草芽胞杆菌BSn5 石油污染 代谢途径 功能基因 实时荧光PCR |
| 英文关键词: B. subtilis BSn5 oil pollution metabolic pathways functional genes fluorescent RT PCR |
| 基金项目:国家国际科技合作项目（2010DFB23160） |
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| 中文摘要: |
| 石油污染已成为全球最严重的环境问题之一，运用生物学方法治理石油污染逐渐成为研究热点。从大港油田原油样品中筛选到12株能降解原油的菌株，分别对其降解特性进行了研究。其中，被命名为BS的菌株对原油的降解效果最好，降解率高达89.1%，经16S rDNA序列比对，与Bacillus subtilis BSn5的相似性达100%。通过生物信息学的方法，分析菌株B. subtilis BSn5的代谢途径，该菌株中有11个基因参与石油烃类化合物的降解过程。最后利用实时荧光PCR技术，测定在石油降解过程中起重要作用的烷基羟化酶基因（alkB）在菌株BS与标准菌株B. subtilis BSn5中的表达量，结果显示，该基因在菌株BS中的表达量较标准菌株B. subtilis BSn5高约2倍，这可能与菌株BS的生存环境有关。本文为检测和分析石油污染，筛选含有降解石油功能基因的微生物及构建工程菌株提供了新的手段。 |
| 英文摘要: |
| Oil pollution has been one of the most serious environment problems the world over and biological methods have become a hot spot for the harnessing the pollutants. In this paper, twelve oil degrading bacterial strains were isolated from Dagang Oil Field in Tianjin, North China and studied their degradation characteristics. Among them a strain named BS showed the highest degradation effect with as high as 89.1%. 16S rDNA sequence comparison showed that it had 100% similarity with Bacillus subtilis BSn5. Metabolic pathways of the strain B. stubilis BSn5 and found through bioinformatics methods that there were about 11 genes involve in oil degradation process. Finally fluorescent RT PCR was used to examine the expression of alkyl hydroxylase B (alkB) gene in BS strain and in standard B. subtilis BSn5. The result showed that the expression in BS strain was two folds higher than those in standard B. subtilis BSn5. It may be related to the habitat environment of BS strain. This study provides a new measure to monitor and analyze the oil pollution, to screen microbes containing functional oil degradation genes, and even to construct the engineering strains to decompose oil. |
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