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采用苯酚羟化酶基因特异引物检测苯酚降解菌
引用本文:徐玉泉,方宣钧,陈明,张维,李浚明,林敏.采用苯酚羟化酶基因特异引物检测苯酚降解菌[J].微生物学报,2001,41(3):298-303.
作者姓名:徐玉泉  方宣钧  陈明  张维  李浚明  林敏
作者单位:1. 中国农业科学院生物技术研究所;中国农业大学生物学院;中国农业科学院原子能利用研究所
2. 中国农业科学院生物技术研究所
3. 中国农业科学院原子能利用研究所
4. 中国农业大学生物学院
基金项目:863计划资助项目(863-SZ-03-01)
摘    要:根据苯酚羟化酶基因高度保守序列设计了一对该基因的特异PCR引物。采用该特异引物从苯酚降解菌醋酸钙不动杆菌 (Acinetobactercalcoaceticus)PHEA 2的总DNA中扩增到唯一一条大小为 684bp的片段。该DNA片段与已知的A .calcoaceticusNCIB82 50的苯酚羟化酶基因具有高度的同源性 ,其核苷酸序列的同源性为 84% ,推导的氨基酸序列的同源性为 98%。对苯酚和非苯酚降解菌株的PCR扩增结果表明 :所有苯酚降解菌均能扩增出 684bp的特征片段 ,而非苯酚降解菌则无PCR条带。对炼焦废水中的细菌群落进行PCR扩增和生化特性检测表明 :显示 684bp特征片段的菌株均具有苯酚降解特性。上述结果表明 ,利用苯酚羟化酶基因的特异引物可对环境中的苯酚降解菌株进行准确快速的PCR检测。

关 键 词:苯酚降解菌    苯酚羟化酶基因    PCR检测    炼焦废水
文章编号:0001-6209(2001)03-0298-06
修稿时间:2000年7月19日

THE DETECTION OF PHENOL DEGRADING STRAIN IN ENVIRONMENT WITH SPECIFIC PRIMER OF PHENOL HYDROXYLASE GENE
Y Xu,X Fang,M Chen,W Zhang,J Li,M Lin.THE DETECTION OF PHENOL DEGRADING STRAIN IN ENVIRONMENT WITH SPECIFIC PRIMER OF PHENOL HYDROXYLASE GENE[J].Acta Microbiologica Sinica,2001,41(3):298-303.
Authors:Y Xu  X Fang  M Chen  W Zhang  J Li  M Lin
Affiliation:College of Biology, China Agricultural University, Institute of Biotechnology Research, Institute for Application of Atomic Energy, CAAS, Beijing, China.
Abstract:A 684 bp oligonucleotide fragment was produced by PCR amplification from phenol-degrading strain Acinetobacter calcoaceticus PHEA-2 with the specific primers of gene encoding phenol hydroxylase. The nucleotide sequence of this fragment and its deduced amino acid sequence share 84% and 98% homology with the phenol hydroxylase gene and its deduced amino acid sequences of phenol-degrading strain Acinetobacter calcoaceticus NCIB8250. Sets of different aromatic compounds degrading strains were used to test this specific prime. The 684 bp-fragments were amplified only from phenol-degrading strains by PCR. When using this pair of primers to detect the bacterial isolates from wastewater discharged from coking plant, all the tested strains, which possess 684 bp characteristic fragment, showed the ability to degrade phenol in this study.
Keywords:Phenol\|degrading strains  Phenol Hydroxylase gene  PCR detection  Coking wastewater
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