采用苯酚羟化酶基因特异引物检测苯酚降解菌  

THE DETECTION OF PHENOL DEGRADING STRAIN IN ENVIRONMENT WITH SPECIFIC PRIMER OF PHENOL HYDROXYLASE GENE 

作　　者：徐玉泉[1] 方宣钧[2] 陈明[3] 张维[3] 李浚明[1] 林敏[3] 

机构地区：[1]中国农业大学生物学院,北京100094 [2]中国农业科学院生物技术研究所,北京100081 [3]中国农业科学院原子能利用研究所,北京100094

出　　处：《微生物学报》2001年第3期298-303,共6页Acta Microbiologica Sinica

基　　金：863计划资助项目 ( 863 SZ 0 3 0 1 )&&

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。A 684bp 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 684bp\|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 684bp characteristic fragment,showed the ability to degrade phenol in this study.

关 键 词：苯酚降解菌 苯酚羟化酶基因 PCR检测 炼焦废水 引物 

分 类 号：Q789[生物学—分子生物学]