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首页> 外文期刊>Environmental Science & Technology >Direct Quantification of Inorganic Polyphosphate in Microbial Cells Using 4'-6-Diamidino-2-Phenylindole (DAPI)
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Direct Quantification of Inorganic Polyphosphate in Microbial Cells Using 4'-6-Diamidino-2-Phenylindole (DAPI)

机译：使用4'-6-Diamidino-2-Phenylindole（DAPI）对微生物细胞中的无机多磷酸盐进行直接定量

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摘要

Inorganic polyphosphate (polyP) is increasingly being recognized as an important phosphorus sink within the environment, playing a central role in phosphorus exchange and phosphogenesis. Yet despite the significant advances made in polyP research there is a lack of rapid and efficient analytical approaches for the quantification of polyP accumulation in microbial cultures and environmental samples. A major drawback is the need to extract polyP from cells prior to analysis. Due to extraction inefficiencies this can lead to an underestimation of both intracellular polyP levels and its environmental pool size; we observed 23—58% loss of polyP using standard solutions and current protocols. Here we report a direct fluorescence based DAPI assay system which removes the requirement for prior polyP extraction before quantification. This increased the efficiency of polyP detection by 28—55% in microbial cultures suggesting quantitative measurement of the intracellular polyP pool. It provides a direct polyP assay which combines quantification capability with technical simplicity. This is an important step forward in our ability to explore the role of polyP in cellular biology and biogeochemical nutrient cycling.
机译：无机多磷酸盐（polyP）日益被认为是环境中重要的磷汇，在磷交换和磷发生中起着核心作用。尽管polyP研究取得了重大进展，但仍缺乏快速有效的分析方法来定量微生物培养物和环境样品中polyP的积累。一个主要缺点是需要在分析之前从细胞中提取polyP。由于提取效率低下，这可能会导致细胞内多聚磷水平及其环境库大小的低估。我们观察到使用标准溶液和当前方案的polyP损失了23-58％。在这里，我们报告了一种基于荧光的直接DAPI分析系统，该系统消除了定量之前需要进行polyP提取的要求。这在微生物培养物中将polyP的检测效率提高了28-55％，表明对细胞内polyP库的定量测量。它提供了一种直接的polyP测定方法，该方法结合了定量功能和技术简便性。这是我们探索polyP在细胞生物学和生物地球化学养分循环中作用的重要一步。

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  • 来源
    《Environmental Science & Technology》 |2011年第18期|p.7799-7803|共5页
  • 作者

    Anna N. Kulakova; Darragh Hobbs; Matthew Smithen; Evgeny Pavlov; Jack A.Gilbert; John P. Quinn; John W. McGrath;

  • 作者单位

    School of Biological Sciences and QUESTOR Centre, The Queen's University of Belfast, Medical Biology Centre, 97 Lisburn Rd.,Belfast, BT9 7BL, Northern Ireland;

    School of Biological Sciences and QUESTOR Centre, The Queen's University of Belfast, Medical Biology Centre, 97 Lisburn Rd.,Belfast, BT9 7BL, Northern Ireland;

    Dalhousie University, Sir Charles Tupper Medical Building, Halifax, Nova Scotia, Canada, B3H 1XSH.;

    Dalhousie University, Sir Charles Tupper Medical Building, Halifax, Nova Scotia, Canada, B3H 1XSH.;

    Argonne National Laboratory, 9700 South Cass Avenue, Argonne, IlUnois 60439, United States Department of Ecology and Evolution, University of Chicago, 5640 South Ellis Avenue, Chicago, Illinois 60637, United States;

    School of Biological Sciences and QUESTOR Centre, The Queen's University of Belfast, Medical Biology Centre, 97 Lisburn Rd.,Belfast, BT9 7BL, Northern Ireland;

    School of Biological Sciences and QUESTOR Centre, The Queen's University of Belfast, Medical Biology Centre, 97 Lisburn Rd.,Belfast, BT9 7BL, Northern Ireland;

  • 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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