Development of an oligonucleotide microarray to detect di- and monooxygenase genes for benzene degradation in soil.
Development of an oligonucleotide microarray to detect di- and monooxygenase genes for benzene degradation in soil.
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DOI：
10.1111/j.1574-6968.2008.01223.x
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发表时间：
2008-08
影响因子：
2.1
通讯作者：
S. Iwai;F. Kurisu;H. Urakawa;O. Yagi;I. Kasuga;H. Furumai
中科院分区：
文献类型：
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作者：
S. Iwai;F. Kurisu;H. Urakawa;O. Yagi;I. Kasuga;H. Furumai
近年来，各种环境基因被发现。为了表征它们的功能，有必要了解哪些基因以及这些基因的组合负责土壤污染物的生物降解。在这篇文章中，一个60聚体寡核苷酸芯片的构建，同时检测苯和相关化合物的双和单加氧酶基因。总共设计了148个探针，并使用以下标准通过纯培养杂交来验证，以区分高度同源的基因：与非靶序列具有≤ 53-bp的同一性和≤ 25-bp的连续延伸。微阵列杂交进行PCR扩增产物从五个苯修正的土壤和两个石油污染的土壤。六个探针给出了一个积极的信号，超过六个土壤，因此，他们可能代表苯在环境中降解的关键序列。本研究建立的基因芯片将为筛选参与苯降解的关键基因和快速分析污染土壤中苯加氧酶基因多样性提供有力的工具。
Diverse environmental genes have been identified recently. To characterize their functions, it is necessary to understand which genes and what combinations of those genes are responsible for the biodegradation of soil contaminants. In this article, a 60-mer oligonucleotide microarray was constructed to simultaneously detect di- and monooxygenase genes for benzene and related compounds. In total, 148 probes were designed and validated by pure-culture hybridizations using the following criteria to discriminate between highly homologous genes: < or =53-bp identities and < or =25-bp continuous stretch to nontarget sequences. Microarray hybridizations were performed using PCR products amplified from five benzene-amended soils and two oil-contaminated soils. Six of the probes gave a positive signal for more than six soils; thus, they may represent key sequences for benzene degradation in the environment. The microarray developed in this study will be a powerful tool for the screening of key genes involved in benzene degradation and for the rapid profiling of benzene oxygenase gene diversity in contaminated soils.