Creating a mutant luciferase resistant to HPV chemical inhibition by random mutagenesis and colony-level screening

Luminescence : the Journal of Biological and Chemical Luminescence
Eileen Kim-ChoiIrina Rybina

Abstract

Firefly luciferase covers a wide range of applications. One common usage of the bioluminescence assay is the measurement of intracellular concentration of adenosine triphosphate (ATP) for cell viability. However, inhibition of the enzyme reaction by chemicals in the assay has so far limited the application of luciferase for high production volume (HPV) chemical testing. The objective of this research was to obtain a mutant luciferase with increased stability to inhibition by HPV chemicals, yet retaining specific activity comparable to, or better than, wild-type luciferase. The enzymatic properties of the wild-type luciferase were improved by random mutagenesis and colony-level screening. In this paper, the detailed process of creating mutant luciferases for testing the toxicity of HPV chemicals is described. As a result, two mutant luciferases were created, with different degrees of improved tolerance to inhibition by chloroform and other HPV chemicals.

References

Jul 1, 1989·Journal of Bioluminescence and Chemiluminescence·K V WoodW D McElroy
Feb 26, 1974·Biochemistry·M DeLuca, W D McElroy
Aug 1, 1997·The Journal of Biological Chemistry·B R BranchiniM H Murtiashaw
May 1, 1960·Archives of Biochemistry and Biophysics·H H SELIGER, W D McELROY
Jul 1, 1993·Toxicology in Vitro : an International Journal Published in Association with BIBRA·S WakuriH Ono

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Citations

Jul 27, 2010·Journal of Photochemistry and Photobiology. B, Biology·João M M Leitão, Joaquim C G Esteves da Silva
Sep 12, 2006·Toxicology in Vitro : an International Journal Published in Association with BIBRA·Eileen Kim-ChoiIrina Rybina
Jan 1, 2012·Computational and Structural Biotechnology Journal·Mikhail I Koksharov, Natalia N Ugarova

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