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Master Thesis

Modification génétique des facteurs de transcription chez Phaeodactylum tricornutum

Abstract : Microalgae are organisms of interest for biofuel production because under stress conditions they produce triglycerides that can be transformed into biodiesel or biokerosene. However, producing biofuels from microalgae is still a laboratory experiment, particularly because of the excessively high production costs. My internship is part of a research project that consists of genetically modifying the microalgae Phaeodactylum tricornutum. This includes modifying it by over-expressing, underexpressing or inactivating genes encoding transcription factors (TFs) potentially involved in lipid production. The purpose of this internship was to: 1) select TFs with marked differential expression under nitrogen stress conditions; 2) construct recombinant plasmids for these candidates using different genetic recombination methods (overexpression, silencing and knock-out (KO)); 3) perform transformations by electroporation and bacterial conjugation. Analysis of the RNA sequencing data revealed 7 candidates from 5 families (aureochrome, CCCH, HSF, MYB and TAZ). Two plasmid constructions for over- and under-expression methods have been successfully realized and the microalgae transformed by electroporation. The results are not yet available. Plasmids for the KO method were constructed by "Golden Gate" and were used in the conjugation transformation technique. The results indicate the presence of zeocin-resistant Pt colonies on petri dishes, confirming the presence of the recombinant plasmid PtPuc3-diaCas9 in the microalgae. Biochemical analyses of mutants under different conditions are now required.
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Master Thesis
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Submitted on : Tuesday, November 19, 2019 - 4:27:41 PM
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Romain Perdoux. Modification génétique des facteurs de transcription chez Phaeodactylum tricornutum. Sciences du Vivant [q-bio]. 2019. ⟨dumas-02370974⟩

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