Microplate Luminometry for Toxicity Bioassay of Chemicals on Luciferase

文献类型： 外文期刊

第一作者： Ge Hui-lin

作者： Ge Hui-lin;Luo Jin-hui;Lu Dai-zhu;Su Bing-xia;Ge Hui-lin;Liu Shu-shen;Chen Fu

作者机构：

关键词： Luciferase;Microplate luminometry;Least square;Dose-response curve;ATP biphasic response

期刊名称：SPECTROSCOPY AND SPECTRAL ANALYSIS （ 影响因子：0.589； 五年影响因子：0.504 ）

ISSN： 1000-0593

年卷期： 2013 年 33 卷 10 期

页码：

收录情况： SCI

摘要： A new microplate luminometry for the toxicity bioassay of chemicals on firefly luciferase, was developed using the multifunctional microplate reader (SpectraMax M5) to measure the luminous intensity of luciferase. Efects of luciferase concentration, luciferin concentration, ATP concentration, pH, temperature, and reaction time on the luminescence were systematically investigated. It was found that ATP exerted a biphasic response on the luciferase luminescence and the maximum relative light units (RLU) occurred at an ATP concentration of 1. 1 X 10(-4) mol . L-1. The method was successfully employed in the toxic effect test of NaF, NaCl, KBr and NaBF4 on luciferase. Using nonlinear least square technique, the dose-response curves (DRC) of the 4 chemicals were accurately fitted with the coefficient of determination (R-2) between the fitted and observed responses being greater than 0. 99. The median effective concentration (EC50) of the 4 chemicals were accurately measured from the DRC models. Compared with some literatures, the bioassay is a fast easy-operate and cost-effective method with high accuracy.

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