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首页> 外文期刊>Environmental Science & Technology >Degradation of Extracellular Antibiotic Resistance Genes with UV_(254) Treatment
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Degradation of Extracellular Antibiotic Resistance Genes with UV_(254) Treatment

机译：UV_（254）处理降解细胞外抗生素抗性基因

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摘要

Disinfected wastewater effluent contains a complex mixture of biomolecules including DNA. If intact genes conveying antibiotic resistance survive the disinfection process, environmental bacteria may take them up. We treated plasmid pWH1266, which contains ampicillin resistance gene bla_(TEM-1) and tetracycline resistance gene tetA, with UV_(254) doses up to 430 mJ/cm~2 and studied the ability of those genes to be acquired by Acinetobacter baylyi. The plasmids required approximately 20-25 mJ/cm~2 per log_(10) loss of transformation efficiency. We monitored plasmid DNA degradation using gel electrophoresis and qPCR with both short amplicons (~200 bps, representative of ARG amplicon lengths commonly used for environmental monitoring) and long amplicons (800—1200 bps, designed to cover the entire resistance genes). The rate of transformability loss due to UV_(254) treatment was approximately 20× and 2× larger than the rate of gene degradation measured with the short and long amplicons qPCR, respectively. When extrapolated to account for the length of the entire pWH1266 plasmid, the qPCR rate constants were 2—7× larger than the rate constants measured with transformation assays. Gel electrophoresis results confirmed that DNA cleavage was not a major inactivating mechanism. Overall, our results demonstrate that qPCR conservatively measures the potential for a gene to be transformed by environmental bacteria following UV_(254) treatment.
机译：消毒后的废水含有复杂的生物分子混合物，包括DNA。如果传达抗生素抗性的完整基因在消毒过程中仍然存在，则环境细菌可能会将其吸收。我们处理了质粒pWH1266，该质粒含有氨苄青霉素抗性基因bla_（TEM-1）和四环素抗性基因tetA，其UV_（254）剂量高达430 mJ / cm〜2，并研究了这些基因被拜氏不动杆菌获得的能力。每转化损失log_（10），质粒需要大约20-25 mJ / cm〜2。我们使用凝胶电泳和qPCR监测了质粒DNA的降解，包括短扩增子（〜200 bps，代表通常用于环境监测的ARG扩增子的长度）和长扩增子（800-1200 bps，旨在覆盖整个抗性基因）。由于使用UV_（254）处理而导致的可转化性丧失速率分别比使用短扩增子和长扩增子qPCR测得的基因降解速率分别大20倍和2倍。当推断出整个pWH1266质粒的长度时，qPCR速率常数比用转化测定法测得的速率常数大2-7倍。凝胶电泳结果证实，DNA切割不是主要的失活机制。总体而言，我们的结果表明，qPCR保守地测量了经过UV_（254）处理后环境细菌转化基因的潜力。

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  • 来源
    《Environmental Science & Technology》 |2017年第11期|6185-6192|共8页
  • 作者

    Pin Hsuan Chang; Brianna Juhrend; Terese M. Olson; Carl F. Marrs; Krista R. Wigginton;

  • 作者单位

    Department of Civil and Environmental Engineering, University of Michigan, Ann Arbor, Michigan 48109, United States;

    Department of Civil and Environmental Engineering, University of Michigan, Ann Arbor, Michigan 48109, United States;

    Department of Civil and Environmental Engineering, University of Michigan, Ann Arbor, Michigan 48109, United States;

    Epidemiology Department, University of Michigan, 1415 Washington Heights, Ann Arbor, Michigan 48109, United States;

    Department of Civil and Environmental Engineering, University of Michigan, Ann Arbor, Michigan 48109, United States;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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  • 正文语种 eng
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