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Selection OF thermostable mutants OF Phrixotrix hirtus luciferase FOR bioanalytical purposes

Grant number: 13/18449-4
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): November 01, 2013
Effective date (End): October 31, 2014
Field of knowledge:Biological Sciences - Biophysics - Radiology and Photobiology
Principal Investigator:Vadim Viviani
Grantee:Gabriela de Oliveira
Home Institution: Centro de Ciências e Tecnologias para a Sustentabilidade (CCTS). Universidade Federal de São Carlos (UFSCAR). Sorocaba , SP, Brazil

Abstract

Luciferases are enzymes that catalyze the oxidization of molecules knownas luciferins resulting in the production of bioluminescence. Amongluciferases of different organisms, those found in fireflies and otherbeetles have been the focus of many studies and are currently applied formany biotechnological purposes, including bioanalysis of ATP, reportergene assays and cellular bioimaging. The luciferases from beetles arequite sensitivity to high temperatures therefore it is necessary toimprove their thermostability in order to apply them for bioimaging athigh-temperature tissues of mammalians. The luciferase of Phrixotrixrailroadworm is especially interesting for haemoglobin rich tissues and pigmented samples,because is the only natural red emitting luciferase to be cloned.However, it is unstable at higher temperatures of mammalian tissues. Theaim of this study is to engineer, select and characterize thermostablemutants of the unique red emitting Phrixotrix railroadworm luciferasethrough site-directed mutations.