Recherche et caractérisation fonctionnelle de gènes impliqués dans la voie de biosynthèse des furocoumarines chez les citrus par des approches moléculaires, biochimiques et bioinformatiques

Abstract : The aim of this project is to identify and characterize genes involved in the furanocoumarins pathway among Citrus. The approach used in this study was the selection of genes potentially involved in this pathway by BLAST and by comparing their expression levels between individuals producing or not this type of molecules. Then, two approaches have been studied for the purpose of characterizing enzymes by a biochemical approach and the bioinformatics approach. The first selection of candidate genes, made by BLASTing, led to the finding of thirteen P450s and fourteen prenyltransferases. Then, the comparison of the levels of gene expression between oranges (low producer of furanocoumarins) and bergamots and grapefruits (high producers) reduced the number of candidate genes encoding four P450s. The first experiments to lead to the characterization of enzymes by biochemical approach were performed. Bacterial transformations on E. coli have been successful for two genes, Ciclev10018132m and Ciclev10028201m. They will enable the heterologous production of P450s in order to characterize their activity by biochemical approaches. Finally, the modeling of the three-dimensional structure of the enzyme CYP71AZ4 should be used as a basis for the modeling of other P450s and the screening of substrates.
Document type :
Master thesis
File URL :
http://docnum.univ-lorraine.fr/prive/BUS_M_2013_COQUERET_VICTOIRE.pdf
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https://hal.univ-lorraine.fr/hal-01779335
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Submitted on : Thursday, April 26, 2018 - 3:11:33 PM
Last modification on : Friday, September 14, 2018 - 1:13:50 AM

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Victoire Coqueret. Recherche et caractérisation fonctionnelle de gènes impliqués dans la voie de biosynthèse des furocoumarines chez les citrus par des approches moléculaires, biochimiques et bioinformatiques. Sciences de l'environnement. 2013. ⟨hal-01779335⟩

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