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Production De Protéines Recombinantes Production de protéines recombinantes par des plantes carnivores génétiquement transformées : application à Drosera rotundifolia et transfert de la technologie à Nepenthes alata Flore Biteau To cite this version: Flore Biteau. Production de protéines recombinantes par des plantes carnivores génétiquement trans- formées : application à Drosera rotundifolia et transfert de la technologie à Nepenthes alata. Sciences agricoles. Institut National Polytechnique de Lorraine, 2009. Français. NNT : 2009INPL022N. tel-01750583 HAL Id: tel-01750583 https://hal.univ-lorraine.fr/tel-01750583 Submitted on 29 Mar 2018 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. AVERTISSEMENT Ce document est le fruit d'un long travail approuvé par le jury de soutenance et mis à disposition de l'ensemble de la communauté universitaire élargie. Il est soumis à la propriété intellectuelle de l'auteur. Ceci implique une obligation de citation et de référencement lors de l’utilisation de ce document. D'autre part, toute contrefaçon, plagiat, reproduction illicite encourt une poursuite pénale. Contact : [email protected] LIENS Code de la Propriété Intellectuelle. articles L 122. 4 Code de la Propriété Intellectuelle. articles L 335.2- L 335.10 http://www.cfcopies.com/V2/leg/leg_droi.php http://www.culture.gouv.fr/culture/infos-pratiques/droits/protection.htm Ecole Doctorale RP2E UMR INPL(ENSAIA)-INRA 1121 Agronomie et Environnement Plant Advanced Technologies SAS Thèse Présentée pour l’obtention du titre de Docteur de l’INPL En Sciences Agronomiques Par Flore BITEAU Production de protéines recombinantes par des plantes carnivores génétiquement transformées : Application à Drosera rotundifolia et transfert de la technologie à Nepenthes alata THESE CONFIDENTIELLE Soutenue à huis clos le 14 Mai 2009, devant le jury composé de : M. Jean-Louis Goergen Professeur, ENSAIA-INPL, Nancy Président du Jury M. Hubert Schaller Chargé de Recherche, HDR, CNRS-IBMP, Strasbourg Rapporteur Mme Véronique Gomord Chargée de Recherche, HDR, CNRS, Rouen Rapportrice M. Frédéric Bourgaud Professeur, ENSAIA-INPL, Nancy Examinateur M. Eric Gontier Professeur, UPJV, Amiens Examinateur Mme Laurence Gaume Chargée de Recherche, CNRS, Montpellier Examinatrice M. Alain Hehn Maître de conférences, HDR, ENSAIA-INPL Nancy Invité à mon grand-père Albert, à mon parrain Claude 1 2 Remerciements REMERCIEMENTS L’ENSAIA, c’est un peu la maison depuis 6 ans … J’y étais (en tant qu’élève) … et j’y suis restée pendant que mes camarades de classe s’en allaient aux quatre coins du monde. Je pensais en avoir fait le tour… Et pourtant…c’était sans connaître l’univers des laboratoires… Elève ingénieur, on passe dans les couloirs, on vole des regards furtifs dans la salle café, regards empreints de curiosité, de respect et d’inquiétude…Et puis, j’ai franchi la barrière. J’ai rejoint ce monde merveilleux de la blouse blanche® comme le dit Julie ou de la Belouse blanche comme le dirait Eric…(et croyez moi, j’en ai frimé un moment auprès de mes anciens collègues de 3A : « ça y est , j’ai des entrées pour la salle café !! »…). Et j’y ai passé 3 années formidables, riches, et palpitantes. Aussi, je tiens à exprimer ici de façon simple et très sincère, ma reconnaissance chaleureuse à toutes les personnes qui ont joué un rôle important durant ma thèse ou qui m’ont apporté de près ou de loin leur soutien : Je souhaite remercier Madame Véronique Gomord, Chargé de recherche au CNRS de Rouen et Mr Hubert Schaller, Chargé de recherche au CNRS à l’IBMP de Strasbourg, d’avoir accepté d’être les rapporteurs de cette thèse. Merci à Mr Jean-Louis Goergen, Professeur à l’ENSAIA et à Madame Laurence Gaume, Chargée de Recherche au CNRS de Montpellier, d’avoir accepté de juger mon travail. Je remercie, Sylvain Plantureux, directeur du Laboratoire Agronomie et Environnement pendant ces 3 ans. Je remercie bien sûr, Fréderic Bourgaud, mon directeur de thèse, (mais aussi nouveau directeur du Laboratoire Agronomie et Environnement et directeur scientifique de PAT SAS). Merci Fred de m’avoir confié ce projet à mes tout débuts, de m’avoir fait tout simplement confiance. Merci pour ton soutien, tes encouragements au moment où j’en avais le plus besoin, et pour tes conseils éclairés durant ces 3 années. Mes remerciements vont ensuite à mes 2 co-directeurs de thèse : A Alain Hehn … Merci Alain, pour ton aide précieuse, ton suivi, tes conseils plus qu’avisés, ta super disponibilité (je cherche des superlatifs), ton soutien moral (service assuré quasiment 24h/24…mails à l’appui. Tu assures aussi le Service Après Thèse ?), tes corrections éclairs et ton implication dans mon travail… Comme diraient certains, « il est fort 3 Remerciements ce Alain ! », avec un fan club à l’ENSAIA pour preuve (« Hehn Addict », c’est bien ça ?). Que dire d’autre, ou comment le dire ?... Un grand Merci tout simplement. A Eric Gontier, directeur de thèse durant ma première année, et désormais co- directeur. Tu as décidé de t’expatrier à Amiens, à mon grand regret !! Merci Eric pour ton énergie communicative, ton optimisme et ton soutien quotidien durant cette première année. Merci pour tes réunions improbables entre 2 parkings, sur le rebord d’une fenêtre, ou même à Germiny ! Merci aussi de m’avoir donné le goût de la recherche dès ma sortie de l’école. Merci à Jean Paul Fèvre, PDG de Plant Advanced Technologies SAS, de m’avoir confié cette mission, de m’avoir fait confiance, et de m’avoir permis de vivre cette grande aventure qu’est la thèse CIFRE. Merci d’avoir eu cette idée lumineuse de PAT Vendredi®, et d’avoir fait de ma thèse un projet passionnant…Et merci pour cette convivialité et cet esprit d’équipe que tu as instaurés au sein de l’entreprise. Mes remerciements vont ensuite à Sissi Miguel, mon ex stagiaire de Master II, ma collègue, mon amie et désormais ma sœur siamoise !! (hein Bernard ?) ... Sissi, merci d’avoir été mon alliée depuis ton arrivée … et d’être venue partager avec moi la lourde tâche qui nous incombe…. D’abord stagiaire exceptionnelle (la plus extraordinaire que je n’ai jamais eue ; tu pleures un peu là ou je continue ?), puis ingénieur R&D chez PAT SAS sur le projet, tu as largement participé et contribué à ce travail de thèse… Merci tout simplement pour ça. Merci pour ton efficacité, et ton aide précieuse… Merci aussi pour ton soutien moral dans mes moments de doutes, tes séances de coaching et de psychothérapie (merci aussi au bureau d’Etienne…), et aussi d’avoir gardé Moumoute durant les périodes sombres… (Tu ne t’y attendais pas à celle la ?). Et puis, il y a tout le reste, bien sûr, que je ne peux dévoiler ici entièrement…Jude, le drover, John John… cette complicité qui nous unit, et ces regards qui ne nécessitent aucune parole… Mes remerciements vont ensuite à la Team PAT SAS Plantes à traire, mes collègues de bureau : A Benoît : merci pour ton soutien, ta disponibilité, et ta forme olympique matinale quotidienne… et à Jean-Marc : merci pour tes conseils notamment en Culture In Vitro, pour ton investissement dans le projet, et pour nos petites séances de papotage… Merci bien sûr, à tous les doctorants de la thésard vallée et aux stagiaires pour cette ambiance chaleureuse de travail et pour tous ces bons moments partagés … Merci à Dao, Minh, Romain, Petit Seb, µSeb, Vincent, Christelle, Camille, Boris, Emma, Robert, 4 Remerciements Mickaël, Natalia, Lama, Guilhem, Hélène. J’en profite aussi pour remercier la Cath, pour tes éclats de rire mythiques et la joie de te croiser dans les couloirs… Je voudrais remercier en particulier Etienne : le don de code, de logiciel, le prêt de bureau, les poires-chocolat …, merci pour ton soutien. J’ai encore une dette envers toi, qui sera bientôt honorée à condition que mon secret ne soit pas dévoilé… (Kyle ? non ...stp…) Un grand Merci aussi à toi Grand Seb…. Pour ton soutien, ta bonne humeur, tes délires (n’arrête surtout pas, je t’en prie !), et ta complicité durant ces 3 ans… tu as apporté beaucoup de rires et de gaité durant ma thèse… et ce n’est bien sûr pas terminé… Merci à Julie, ma « coloc » de première année…que de bons souvenirs avec toi, Miss M : Shepperd, les belles plantes, Harry, Bob, Christina, ton chili con carne… j’en passe et des meilleurs... Merci à Martine, tu m’as bien souvent facilité la tâche dans mes manips… !! Merci aussi pour ta disponibilité… Merci également à Dominique et à Claude, pour leur aide et leur gestion des phytotrons. Merci à Thamara, pour ta patience d’ange … Merci à Bernard Cunin et à Marc Rémy, pour leur aide et leur disponibilité au Jardin Botanique du Montet. Merci à Jacques Boulay, pour ses conseils de culture des plantes carnivores, et pour m’avoir prêté ses « bibles ». Merci à Emmanuel Guédon, et Céline Chéry pour leur aide et leur disponibilité concernant les expérimentations sur l’interféron gamma et sur le facteur intrinsèque. Merci à Jean-Louis Guéant, pour son implication dans le projet. Enfin, un grand grand grand MERCI à ma famille : Tout d’abord à mes parents, à qui je dois beaucoup. Merci de m’avoir soutenue quotidiennement durant ces 8 années d’études, d’avoir cru en moi, et de m’avoir encouragée pour en arriver là ! ..
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