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用于检测镍、钴和锌的生物发光蓝藻报告菌株的构建。

Construction of bioluminescent cyanobacterial reporter strains for detection of nickel, cobalt and zinc.

作者信息

Peca Loredana, Kós Péter B, Máté Zoltan, Farsang Andrea, Vass Imre

机构信息

Biological Research Center, Institute of Plant Biology, Szeged, Hungary.

出版信息

FEMS Microbiol Lett. 2008 Dec;289(2):258-64. doi: 10.1111/j.1574-6968.2008.01393.x.

Abstract

Two whole-cell bioluminescent reporters were constructed by fusing the reporter genes luxAB with the Co(2+) and Zn(2+) inducible coaT promoter or the Ni(2+)-inducible nrsBACD promoter, respectively, in the genome of Synechocystis sp. PCC 6803. The obtained reporters, designated coaLux and nrsLux, respectively, responded quantitatively to metal ions. After 3 h incubation at 40 micromol m(-2) s(-1) visible light, the detection range of coaLux was 0.3-6 microM for Co(2+) and 1-3 microM for Zn(2+). Incubation in darkness increased the detection range by about four times. The nrsLux reporter was specific to Ni(2+), with a detection range of 0.2-6 microM. However, its activity was inhibited by Zn(2+) with a half maximal inhibitory concentration c. 6 microM, and totally inhibited by darkness. This is the first whole-cell Ni(2+)-specific reporter with a clear dose-signal relationship. In a soil-like mixture of different chemical and oil industry wastes, the coaLux reporter strain detected about 90% of the zinc content of the sample. This study demonstrates the potential for development of a rapid, simple and economical field assay for nickel, cobalt and zinc detection using the coaLux and nrsLux reporters.

摘要

通过将报告基因luxAB分别与集胞藻PCC 6803基因组中Co(2+)和Zn(2+)诱导型coaT启动子或Ni(2+)诱导型nrsBACD启动子融合,构建了两种全细胞生物发光报告基因。所获得的报告基因分别命名为coaLux和nrsLux,它们对金属离子有定量响应。在40 μmol m(-2) s(-1)可见光下孵育3小时后,coaLux对Co(2+)的检测范围为0.3 - 6 μM,对Zn(2+)的检测范围为1 - 3 μM。在黑暗中孵育可使检测范围增加约四倍。nrsLux报告基因对Ni(2+)具有特异性,检测范围为0.2 - 6 μM。然而,其活性受到Zn(2+)的抑制,半数最大抑制浓度约为6 μM,并且在黑暗中完全被抑制。这是首个具有明确剂量 - 信号关系的全细胞Ni(2+)特异性报告基因。在不同化学和石油工业废物的类土壤混合物中,coaLux报告菌株检测到了样品中约90%的锌含量。本研究证明了利用coaLux和nrsLux报告基因开发一种快速、简单且经济的镍、钴和锌检测现场分析方法的潜力。

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