Peca Loredana, Kós Péter B, Máté Zoltan, Farsang Andrea, Vass Imre
Biological Research Center, Institute of Plant Biology, Szeged, Hungary.
FEMS Microbiol Lett. 2008 Dec;289(2):258-64. doi: 10.1111/j.1574-6968.2008.01393.x.
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.
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。