测定化学物对萤光素酶抑制毒性的微板发光法研究

doi:10.3964/j.issn.1000-0593(2013)10-2766-05

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Abstract 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 1.1×10^-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 (R²) 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.

Key words：Luciferase;Microplate luminometry;Least square;Dose-response curve;ATP biphasic response

Received: 2012-12-29 Accepted: 2013-03-06

ZTFLH:

O657.3

Corresponding Authors: LIU Shu-shen E-mail: ssliuhl@263.net

Cite this article:

GE Hui-lin,LIU Shu-shen,CHEN Fu, et al. Microplate Luminometry for Toxicity Bioassay of Chemicals on Luciferase [J]. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2013, 33(10): 2766-2770.

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http://www.gpxygpfx.com/EN/10.3964/j.issn.1000-0593(2013)10-2766-05 OR http://www.gpxygpfx.com/EN/Y2013/V33/I10/2766

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