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Optimization of Bacterial Whole Cell Bioreporters for Toxicity Assay of Environmental Samples

机译：细菌全细胞生物报告物对环境样品毒性测定的优化

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摘要

In a study to optimize bacterial whole cell biosensors (bioreporters) for the detection of environmental contaminants, we constructed a toxicity sensing strain Acinetobacter baylyiADPlrecAJux. The ADP1_recA_lux is a chromosomally based bioreporter which makes the sensing system stable and negates the need for antibiotics to maintain the trait. The ADP1_recAJux is activated to express bioluminescence when it is exposed to DNA damaging toxicants. Since the ADPl_recA_lux constantly expresses a baseline level of bioluminescence, false negative results are avoided. The host strain, A. baylyi ADP1, is an ideal model strain typical of water and soil bacteria occurring in the natural environment, and it is more robust than E. coli in terms of viability, maintenance, and storage. The expression of reporter genes - luxCDABE cloned from Photorhabdus luminescens - is robust in a broad range of temperature (10-40℃). The ADP1_recA_lux was used to detect a variety of toxic or potentially toxic compounds including mitomycin C (MMC), methyl methanesulfonate, ethidium bromide, H_2O_2, toluene, single-wall nanocarbon tubes (SWNCT), nano Au colloids (20 nm), pyrene, beno[a]pyrene, and UV light These exposures revealed that the ADP1_recA_lux was able to detect both genotoxicity and cvtoxicity, qualitatively and quantitatively. The optimal induction time of the ADP1_recA_lux bioreporter was 3 h, and the detection limits for MMC and benezo[a]pyrene were 1.5 nM and 0.4 nM, respectively. The ADPl_recA_lux was also used to detect toxicity of groundwater contaminated by a mixture of phenolic compounds, and the bioreporter toxicity detection was in a good agreement with chemical analysis. The optimized whole cell bioreporter ADPl_recAJux could bernvaluable in providing a simple, rapid, stable, quantitative, robust, and costly efficient approach forthe detection of toxicity in environmental samples.
机译：在一项优化细菌全细胞生物传感器（生物报告者）以检测环境污染物的研究中，我们构建了毒性感应菌株BayineiADPlrecAJux。 ADP1_recA_lux是基于染色体的生物报告基因，可使传感系统稳定并无需使用抗生素来维持该特性。当ADP1_recAJux暴露于DNA破坏性有毒物质时，它被激活以表达生物发光。由于ADP1_recA_lux不断表达生物发光的基线水平，因此可以避免假阴性结果。宿主菌株Baylyi ADP1是理想的模型菌株，典型地存在于自然环境中的水和土壤细菌，并且在活力，维持和储存方面比大肠杆菌更强大。报道基因的表达（从光生光杆菌克隆的luxCDABE）在很宽的温度范围（10-40℃）中都很稳定。 ADP1_recA_lux用于检测多种有毒或潜在有毒化合物，包括丝裂霉素C（MMC），甲磺酸甲酯，溴化乙锭，H_2O_2，甲苯，单壁纳米碳管（SWNCT），纳米金胶体（20 nm），pyr，苯并[a] py和紫外光这些暴露表明，ADP1_recA_lux能够定性和定量检测基因毒性和腔毒性。 ADP1_recA_lux生物报告基因的最佳诱导时间为3 h，MMC和苯并[a] re的检出限分别为1.5 nM和0.4 nM。 ADP1_recA_lux还用于检测被酚类化合物混合物污染的地下水的毒性，生物报告剂毒性检测与化学分析非常吻合。优化的全细胞生物报告基因ADP1_recAJux可以提供一种简单，快速，稳定，定量，鲁棒和昂贵的高效方法来检测环境样品中的毒性。

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  • 来源
    《Environmental Science & Technology》 |2009年第20期|7931-7938|共8页
  • 作者

    YIZHI SONG; GUANGHE LI; STEVEN F. THORNTON; IAN P. THOMPSON; STEVEN A. BANWART; DAVID N. LERNER; WEI E. HUANG;

  • 作者单位

    Department of Environmental Science and Eneineering, Tsinghua University, Beijing, P.R. China Department of Civil and Structural Engineering, Kroto Research Institute, University of Sheffield, Broad Lane, Sheffield S3 7HQ, U.K. State Key Joint Laboratory of Environment Simulation and Pollution Control, Beijing, P.R. China;

    Department of Environmental Science and Eneineering, Tsinghua University, Beijing, P.R. China State Key Joint Laboratory of Environment Simulation and Pollution Control, Beijing, P.R. China;

    Department of Civil and Structural Engineering, Kroto Research Institute, University of Sheffield, Broad Lane, Sheffield S3 7HQ, U.K.;

    Department of Engineering, University of Oxford, U.K.;

    Department of Civil and Structural Engineering, Kroto Research Institute, University of Sheffield, Broad Lane, Sheffield S3 7HQ, U.K.;

    Department of Civil and Structural Engineering, Kroto Research Institute, University of Sheffield, Broad Lane, Sheffield S3 7HQ, U.K.;

    Department of Civil and Structural Engineering, Kroto Research Institute, University of Sheffield, Broad Lane, Sheffield S3 7HQ, U.K.;

  • 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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