{"id":22443,"date":"2003-04-04T00:00:00","date_gmt":"2003-04-04T00:00:00","guid":{"rendered":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/expresion-recombinante-en-e-coli-de-anta%c2%adgenos-de-actinobacillus-pleuropneumoniae-para-vacunacion-y-diagnostico\/"},"modified":"2003-04-04T00:00:00","modified_gmt":"2003-04-04T00:00:00","slug":"expresion-recombinante-en-e-coli-de-anta%c2%adgenos-de-actinobacillus-pleuropneumoniae-para-vacunacion-y-diagnostico","status":"publish","type":"post","link":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/expresion-recombinante-en-e-coli-de-anta%c2%adgenos-de-actinobacillus-pleuropneumoniae-para-vacunacion-y-diagnostico\/","title":{"rendered":"Expresi\u00f3n recombinante en e.coli de ant\u00edgenos de actinobacillus pleuropneumoniae para vacunaci\u00f3n y diagn\u00f3stico"},"content":{"rendered":"<h2>Tesis doctoral de <strong> Andr\u00e9s Medrano Mu\u00f1oz <\/strong><\/h2>\n<p>Actinobacillus pleuropneumoniae es una bacteria gramnegativa que provoca la pleuroneumon\u00eda porcina.  en este trabajo se ha procedido a la producci\u00f3n y purificaci\u00f3n, mediante t\u00e9cnicas de biolog\u00eda molecular, de ant\u00edgenos proteicos de esta bacteria y a su uso en la formulaci\u00f3n de una vacuna por subunidades y de un elisa para diagn\u00f3stico. Los cuatro ant\u00edgenos escogidos fueron dos prote\u00ednas de membrana externa (tbp1 y tbp2) y dos exotoxinas (apxi y apxiii).  para la detecci\u00f3n de tbp1 se utiliz\u00f3 como sonda el gen tbp2. Tras su identificaci\u00f3n, el gen tbp1 fue clonado en el vector puc119 obteni\u00e9ndose su secuencia completa que se deposit\u00f3 en el genbank z49708, habi\u00e9ndose obtenido su patente europea ep0733708 y americana 08\/624655. Finalmente, el resultado fue publicado (daban et al., 1996; medrano et al., 1997) (anexos vii.1 y vii.2). Para la clonaci\u00f3n de apxia y apxiiia, al conocerse sus secuencias, se dise\u00f1aron cebadores de pcr con mutaciones que creaban dianas de restricci\u00f3n, permitiendo as\u00ed la amplificaci\u00f3n sobre el genoma y posterior clonaci\u00f3n en vectores de expresi\u00f3n.  una vez clonados los genes codificantes para las cuatro prote\u00ednas se pas\u00f3 a la producci\u00f3n heter\u00f3loga en escherichia coli. Para ello se utilizaron diversos vectores: pmal, que generan como producto de expresi\u00f3n una prote\u00edna de fusi\u00f3n con la mbp (maltose binding protein) y vectores de la gama pet, con los cuales se ha desarrollado la mayor parte del trabajo. A partir de los extractos obtenidos se procedi\u00f3 a la purificaci\u00f3n de los ant\u00edgenos por cromatograf\u00eda de afinidad aprovechando fusiones con colas de histidinas.  tras producir y purificar los cuatro ant\u00edgenos, se procedi\u00f3 a la elaboraci\u00f3n de una vacuna experimental. Se desarrollaron tres protocolos de vacunaci\u00f3n, estudi\u00e1ndose la inocuidad y efectividad de esa vacuna. Se determin\u00f3 la respuesta humoral a partir de las muestras de suero obtenidas en los ensayos de vacunaci\u00f3n mediante<\/p>\n<p>&nbsp;<\/p>\n<h3>Datos acad\u00e9micos de la tesis doctoral \u00ab<strong>Expresi\u00f3n recombinante en e.coli de ant\u00edgenos de actinobacillus pleuropneumoniae para vacunaci\u00f3n y diagn\u00f3stico<\/strong>\u00ab<\/h3>\n<ul>\n<li><strong>T\u00edtulo de la tesis:<\/strong>\u00a0 Expresi\u00f3n recombinante en e.coli de ant\u00edgenos de actinobacillus pleuropneumoniae para vacunaci\u00f3n y diagn\u00f3stico <\/li>\n<li><strong>Autor:<\/strong>\u00a0 Andr\u00e9s Medrano Mu\u00f1oz <\/li>\n<li><strong>Universidad:<\/strong>\u00a0 Aut\u00f3noma de barcelona<\/li>\n<li><strong>Fecha de lectura de la tesis:<\/strong>\u00a0 04\/04\/2003<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<h3>Direcci\u00f3n y tribunal<\/h3>\n<ul>\n<li><strong>Director de la tesis<\/strong>\n<ul>\n<li>Enrique Querol Murillo<\/li>\n<\/ul>\n<\/li>\n<li><strong>Tribunal<\/strong>\n<ul>\n<li>Presidente del tribunal: francesc xavier Avil\u00e9s puigvert <\/li>\n<li>carles Casas alvero (vocal)<\/li>\n<li> Lloren\u00ed\u00a7 i duran rafel (vocal)<\/li>\n<li>enric Espu\u00f1a mas\u00f3 (vocal)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Tesis doctoral de Andr\u00e9s Medrano Mu\u00f1oz Actinobacillus pleuropneumoniae es una bacteria gramnegativa que provoca la pleuroneumon\u00eda porcina. en este trabajo [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"site-sidebar-layout":"default","site-content-layout":"","ast-site-content-layout":"","site-content-style":"default","site-sidebar-style":"default","ast-global-header-display":"","ast-banner-title-visibility":"","ast-main-header-display":"","ast-hfb-above-header-display":"","ast-hfb-below-header-display":"","ast-hfb-mobile-header-display":"","site-post-title":"","ast-breadcrumbs-content":"","ast-featured-img":"","footer-sml-layout":"","theme-transparent-header-meta":"","adv-header-id-meta":"","stick-header-meta":"","header-above-stick-meta":"","header-main-stick-meta":"","header-below-stick-meta":"","astra-migrate-meta-layouts":"default","ast-page-background-enabled":"default","ast-page-background-meta":{"desktop":{"background-color":"var(--ast-global-color-4)","background-image":"","background-repeat":"repeat","background-position":"center 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