{"id":553,"date":"2012-10-19T15:44:53","date_gmt":"2012-10-19T14:44:53","guid":{"rendered":"http:\/\/saber.comule.com\/?p=553"},"modified":"2020-11-12T19:01:07","modified_gmt":"2020-11-12T18:01:07","slug":"les-vecteurs","status":"publish","type":"post","link":"http:\/\/saber-benmimoun.com\/?p=553","title":{"rendered":"Les vecteurs"},"content":{"rendered":"<p><strong><span style=\"color: #800000;\">I) D\u00e9finition:<\/span><\/strong><br \/>\n<span style=\"color: #333333;\">Petites mol\u00e9cules d&#8217;ADN poss\u00e9dant leur propre syst\u00e8me de r\u00e9plication(origine de replication). Ils servent de v\u00e9hicule<\/span><br \/>\n<span style=\"color: #333333;\"> pour le clonage g\u00e9nique.<\/span><\/p>\n<p align=\"justify\">vecteur natif = vecteur sans insert<\/p>\n<p align=\"justify\">Vecteur recombin\u00e9 = vecteur + insert<\/p>\n<p><strong><span style=\"color: #800000;\">II) Caract\u00e9ristiques des vecteurs:<\/span><\/strong><br \/>\n1. r\u00e9plication \u00e9pisomale (facteur sexuel F):\u00a0r\u00e9plication ind\u00e9pendante de l&#8217;ADN de la cellule h\u00f4te.<br \/>\n2. petite taille:\u00a0pour permettre l&#8217;insertion de grand fragment d&#8217;ADN \u00e9tranger (vecteur\u00a0recombinant qui doit \u00eatre stable)<br \/>\n3. pr\u00e9sence de g\u00e8nes de s\u00e9lection:\u00a0s\u00e9lection des cellules h\u00f4tes qui ont int\u00e9gr\u00e9 un vecteur.<br \/>\n4. pr\u00e9sence de sites de restriction uniques localis\u00e9s dans les g\u00e8nes de s\u00e9lection:\u00a0permet de s\u00e9lectionner les cellules h\u00f4tes qui ont int\u00e9gr\u00e9 un vecteur recombinant.<br \/>\n5. stabilit\u00e9: maintient sans modification dans la cellule h\u00f4te, quel que soit le nombre\u00a0de division (en relation avec la 2.)<\/p>\n<p><strong><span style=\"color: #800000;\"><strong>II<strong>I<\/strong>)\u00a0<\/strong>Les 2 cat\u00e9gories de vecteurs<\/span><\/strong><\/p>\n<p><strong><span style=\"color: #000000;\">Vecteur de clonage<\/span><\/strong><\/p>\n<ul>\n<li>Destin\u00e9 \u00e0 isoler physiquement un fragment d&#8217;ADN et \u00e0 amplifier le nombre\u00a0de copies.<\/li>\n<li>Poss\u00e8de un polylinker(appel\u00e9 aussi\u00a0<em>multiple cloning site CMS)<\/em><\/li>\n<\/ul>\n<p><strong><span style=\"color: #000000;\">Vecteur d&#8217;expression<\/span><\/strong><\/p>\n<ul>\n<li><span style=\"color: #333333;\"><strong style=\"color: #000000;\">\u00a0<\/strong>maintient et expression d&#8217;un nsert<\/span><\/li>\n<li>promoteur constitutif ou inductible (IPTG, Arabinose) compatible avec l&#8217;hote<\/li>\n<li>terminateur de la transcription<\/li>\n<li>La s\u00e9quence SD=Shine Dalgarno (RBS chez les bact\u00e9ries)<\/li>\n<li>optionnel : His Tag pour faciliter la purification de la future proteine.<\/li>\n<\/ul>\n<p><span style=\"color: #800000;\"><strong>IV) Diff\u00e9rents vecteurs de clonage:<\/strong><\/span><\/p>\n<p><span style=\"color: #008000;\"><strong>1. Les plasmides:<\/strong><\/span><\/p>\n<div id=\"attachment_598\" style=\"width: 160px\" class=\"wp-caption alignright\"><a href=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/10\/pic001.jpg\"><img loading=\"lazy\" decoding=\"async\" aria-describedby=\"caption-attachment-598\" class=\"size-thumbnail wp-image-598\" title=\"\" src=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/10\/pic001-150x150.jpg\" alt=\"\" width=\"150\" height=\"150\" \/><\/a><p id=\"caption-attachment-598\" class=\"wp-caption-text\">Exemple de plasmide<\/p><\/div>\n<p>C&#8217;est une petite mol\u00e9cule d&#8217;ADN circulaire poss\u00e9dant son propre syst\u00e8me de r\u00e9plication. Il est d&#8217;origine bact\u00e9rienne. On peut obtenir un plasmide navette qui peut etre utilis\u00e9 chez la levure Saccharomyces cerevisiae, en ajoutant une origine de replication 2\u03bc pour la levure, et un facteur de s\u00e9lection comme l&#8217;auxotrophie pour His.<br \/>\nCaract\u00e9ristiques des plasmides:<\/p>\n<ul>\n<li>Taille max de l&#8217;insert = 20 Kb<\/li>\n<li>Peuvent \u00eatre pr\u00e9sent en plusieurs centaines de copies au sein d&#8217;une m\u00eame cellule bact\u00e9rienne (cela d\u00e9pend de la souche bact\u00e9rienne).<\/li>\n<li>Pr\u00e9sence de la s\u00e9quence ORI (qui permet la r\u00e9plication autonome)<\/li>\n<li>Petite taille: 2 \u00e0 5 kb (le chromosome d&#8217;E. coli fait 4700kb)<\/li>\n<li>G\u00e8nes port\u00e9s: g\u00e8nes de r\u00e9sistance aux antibiotiques (g\u00e8nes de s\u00e9lection), g\u00e8nes codant pour des toxines et des antibiotiques et des g\u00e8nes codant pour la d\u00e9gradation de certains compos\u00e9s aromatiques.<\/li>\n<li>contient des sites de restriction uniques \u00e0 l&#8217;ext\u00e9rieur et \u00e0 l&#8217;int\u00e9rieur des g\u00e8nes de s\u00e9lection.<\/li>\n<\/ul>\n<p><span style=\"color: #008000;\"><strong>2. Les vecteurs phagiques:<br \/>\n<\/strong><strong><a href=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/10\/pic004.jpg\"><img loading=\"lazy\" decoding=\"async\" class=\"alignright size-thumbnail wp-image-609\" title=\"\" src=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/10\/pic004-150x150.jpg\" alt=\"\" width=\"150\" height=\"150\" \/><\/a><\/strong><\/span><\/p>\n<ul>\n<li>Taille max de l&#8217;insert = 25 Kb \u00a0(correspond au remplacement de la r\u00e9gion b2 non<\/li>\n<li>essentielle)<\/li>\n<li>Entr\u00e9e par transduction ou infection, spontan\u00e9es des cellules bact\u00e9riennes.<\/li>\n<li>Extr\u00e9mit\u00e9s Cos permettant sa recircularisation \u00e0\u00a0l\u2019int\u00e9rieur\u00a0de\u00a0l\u2019h\u00f4te.<\/li>\n<li>permet de construire une banque \u00a0d&#8217;ADN g\u00e9nomique.<\/li>\n<\/ul>\n<div><\/div>\n<div><\/div>\n<div><\/div>\n<div><span style=\"color: #008000;\"><strong>3. Cosmides<\/strong><\/span><\/div>\n<p><strong><a href=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/10\/pic009.gif\"><img loading=\"lazy\" decoding=\"async\" class=\"size-thumbnail wp-image-584 alignright\" title=\"\" src=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/10\/pic009-150x150.gif\" alt=\"\" width=\"150\" height=\"150\" \/><\/a><\/strong>C&#8217;est un\u00a0hybrides entre les plasmides et le bact\u00e9riophage\u00a0<img loading=\"lazy\" decoding=\"async\" src=\"http:\/\/www.edu.upmc.fr\/sdv\/masselot_05001\/images\/lambda.gif\" alt=\"\" width=\"8\" height=\"11\" \/>.<\/p>\n<div>\n<ul>\n<li>Taille max de l&#8217;insert = 50Kb<\/li>\n<li>Origine de r\u00e9plication<\/li>\n<li>Un polylinker<\/li>\n<li>Un g\u00e8ne conf\u00e9rant un r\u00e9sistance \u00e0 un antibiotique<\/li>\n<li>les extr\u00e9mit\u00e9s cos du phage <img loading=\"lazy\" decoding=\"async\" src=\"http:\/\/www.edu.upmc.fr\/sdv\/masselot_05001\/images\/lambda.gif\" alt=\"\" width=\"8\" height=\"11\" \/>,\u00a0et donc l&#8217;aptitude \u00e0 \u00eatre recircularis\u00e9\u00a0apr\u00e8s\u00a0l\u2019entr\u00e9e\u00a0dans\u00a0l\u2019h\u00f4te.<\/li>\n<li>permet la\u00a0cr\u00e9ation\u00a0de banque g\u00e9nomique<\/li>\n<\/ul>\n<\/div>\n<div id=\"attachment_596\" style=\"width: 160px\" class=\"wp-caption alignright\"><a href=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/10\/pic012.gif\"><img loading=\"lazy\" decoding=\"async\" aria-describedby=\"caption-attachment-596\" class=\"size-thumbnail wp-image-596   \" style=\"border-style: none; margin: 0px; padding: 0px; -webkit-user-drag: none;\" src=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/10\/pic012-150x150.gif\" alt=\"\" width=\"150\" height=\"150\" srcset=\"http:\/\/saber-benmimoun.com\/wp-content\/uploads\/2012\/10\/pic012-150x150.gif 150w, http:\/\/saber-benmimoun.com\/wp-content\/uploads\/2012\/10\/pic012-300x300.gif 300w, http:\/\/saber-benmimoun.com\/wp-content\/uploads\/2012\/10\/pic012.gif 305w\" sizes=\"auto, (max-width: 150px) 100vw, 150px\" \/><\/a><p id=\"caption-attachment-596\" class=\"wp-caption-text\">Exemple de BAC<\/p><\/div>\n<div><strong><span style=\"color: #008000;\">4. BAC<\/span>\u00a0<\/strong>(Bacterial Artificiel Chromosome)<\/p>\n<ul>\n<li>Taille max de l&#8217;insert = 300 Kb<\/li>\n<li>ADN circulaire<\/li>\n<li>Origine de r\u00e9plication issue de l&#8217;\u00e9pisome sexuel F de E. coli.<\/li>\n<li>Faible nombre de copies<\/li>\n<li>Marqueur de s\u00e9lection<\/li>\n<li>Entr\u00e9e par electroporation<\/li>\n<li>peu d&#8217;\u00e9v\u00e8nement de recombinaison avec l&#8217;ADNg de\u00a0l\u2019h\u00f4te.<\/li>\n<\/ul>\n<p><strong><span style=\"color: #008000;\">5. YAK<\/span>\u00a0<\/strong>(Yeast Artificial Chromosome)<\/p>\n<div id=\"attachment_594\" style=\"width: 160px\" class=\"wp-caption alignright\"><a href=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/10\/fig_2_22.gif\"><img loading=\"lazy\" decoding=\"async\" aria-describedby=\"caption-attachment-594\" class=\"size-thumbnail wp-image-594 \" title=\"YAK\" src=\"http:\/\/saber.comule.com\/wp-content\/uploads\/2012\/10\/fig_2_22-150x138.gif\" alt=\"\" width=\"150\" height=\"138\" \/><\/a><p id=\"caption-attachment-594\" class=\"wp-caption-text\">YAK<\/p><\/div>\n<ul>\n<li>Taille max de l&#8217;insert = 1500 Kb<\/li>\n<li>un ou plusieurs marqueurs de s\u00e9lection<\/li>\n<li>ADN lin\u00e9aire<\/li>\n<li>Centrom\u00e8re (CEN) et t\u00e9lom\u00e8res.<\/li>\n<li>Origine de replication pour la levure: Autonomous Replication Site (ARS)<\/li>\n<li>Polylinker (=MCS)<\/li>\n<li>Entr\u00e9e par transformation<\/li>\n<li>tr\u00e8s instable car forte fr\u00e9quence de recombinaison homologue avec l&#8217;ADNg<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<\/div>\n","protected":false},"excerpt":{"rendered":"<p>I) D\u00e9finition: Petites mol\u00e9cules d&#8217;ADN poss\u00e9dant leur propre syst\u00e8me de r\u00e9plication(origine de replication). Ils servent de v\u00e9hicule pour le clonage g\u00e9nique. vecteur natif = vecteur sans insert Vecteur recombin\u00e9 = vecteur + insert II) Caract\u00e9ristiques des vecteurs: 1. r\u00e9plication \u00e9pisomale (facteur sexuel F):\u00a0r\u00e9plication ind\u00e9pendante de l&#8217;ADN de la cellule h\u00f4te. 2. petite taille:\u00a0pour permettre l&#8217;insertion&hellip;<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":[],"categories":[7],"tags":[],"class_list":["post-553","post","type-post","status-publish","format-standard","hentry","category-biologie","category-7","description-off"],"_links":{"self":[{"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=\/wp\/v2\/posts\/553","targetHints":{"allow":["GET"]}}],"collection":[{"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=553"}],"version-history":[{"count":59,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=\/wp\/v2\/posts\/553\/revisions"}],"predecessor-version":[{"id":2958,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=\/wp\/v2\/posts\/553\/revisions\/2958"}],"wp:attachment":[{"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=553"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=553"},{"taxonomy":"post_tag","embeddable":true,"href":"http:\/\/saber-benmimoun.com\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=553"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}