Diverse aadA gene cassettes on class 1 integrons introduced into soil via spread manure

Diverse aadA gene cassettes on class 1 integrons introduced into soil via spread manure
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DOI:
10.1016/j.resmic.2009.06.005
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发表时间:
2009-07-01
影响因子:
2.6
通讯作者:
Smalla, Kornelia
Smalla, Kornelia
中科院分区:
生物学3区
文献类型:
--
作者:
Binh, Chu T. T.;Heuer, Holger;Smalla, Kornelia

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1类整合子被认为在抗生素耐药基因的传播中起重要作用。临床分离的1类整合子通常在基因盒整合的可变区域两侧具有5‘和3’保守片段。利用锁定保守片段的引物从粪便和土壤的总群落DNA中扩增可变区。采用克隆、限制性片段长度多态性(RFLP)分析和测序等方法研究了粪便和肥料土壤中整合子基因盒的多样性。所分析的整合子组可以在施用过肥料的土壤中检测到,而在没有施用过肥料的土壤中检测不到。在克隆文库中经常发现编码链霉素和大观霉素耐药的aadA基因盒。为了阐明“临床”1类整合子基因盒是否通常通过粪便引入农业土壤,对来自15个不同养猪场的一组独特粪便样本进行了调查。在所有农田尺度的粪便中都检测到aadA基因盒,证实了定位于I类整合子上的aadA基因是通过猪粪进入农业土壤的。此外,在许多肥料的总群落DNA中检测到不带基因盒的I类整合子。(C) 2009 Elsevier Masson SAS。版权所有。
Class 1 integrons are assumed to play an important role in dissemination of antibiotic resistance genes. Class 1 integrons from clinical isolates often have 5'- and 3'-conserved segments flanking the variable region where gene cassettes integrate. Primers targeting the conserved segments were used to amplify the variable region from total community DNA of manure and soil. Integron gene cassette diversity in manure and manured soils was studied by cloning, restriction fragment length polymorphism (RFLP) analysis and sequencing. The analyzed group of integrons could be detected in manured soils but not in soil without a history of manure treatment. aadA gene cassettes encoding resistance to streptomycin and spectinomycin were frequently detected in the clone libraries. To elucidate whether "clinical" class 1 integron gene cassettes are typically introduced via manure into agricultural soil, a survey with a unique set of manure samples originating from 15 different pig-producing facilities was performed. aadA gene cassettes were detected in all field-scale manures, confirming that aadA genes localized on class I integrons are introduced via piggery manure into agricultural soils. In addition, class I integrons without gene cassettes were detected in total community DNA of many manures. (C) 2009 Elsevier Masson SAS. All rights reserved.