{"id":86834,"date":"2000-06-10T00:00:00","date_gmt":"2000-06-10T00:00:00","guid":{"rendered":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/construccion-de-un-cdna-infectivo-del-coronavirus-de-la-gastorenteritis-porcina-transmisible\/"},"modified":"2000-06-10T00:00:00","modified_gmt":"2000-06-10T00:00:00","slug":"construccion-de-un-cdna-infectivo-del-coronavirus-de-la-gastorenteritis-porcina-transmisible","status":"publish","type":"post","link":"https:\/\/www.deberes.net\/tesis\/quimica\/construccion-de-un-cdna-infectivo-del-coronavirus-de-la-gastorenteritis-porcina-transmisible\/","title":{"rendered":"Construccion de un cdna infectivo del coronavirus de la gastorenteritis porcina transmisible"},"content":{"rendered":"<h2>Tesis doctoral de <strong>  Gonzalez Martienz Jos\u00e9 Manuel <\/strong><\/h2>\n<p>El objetivo final de este trabajo ha sido la obtenci\u00f3n de un cdna infectivo correspondiente al genoma del coronavirus de la gastroenteritis porcina transmisible (vgpt), que permita realizar realizar genetica inversa con el genoma viral para estudiar los mecanismos de replicacion y transcripcion y su uso como vector viral. Los objetivos parciales llevados a cabo han sido:  1. Estudio de la capacidad autorreplicativa del genoma defectivo di-a, que contiene el gen la replicasa viral. Se ha demostrado que el genoma defectivo di-a, derivado del vgpt, es un rna que se replica autonomamente sin el requerimiento de un virus complementador.  2. Clonaje del rna autorreplicativo en un pl\u00e1smido procariotico utilizando estrategias que evitan la toxicidad de la secuencia viral en las celulas procarioticos. Se ha obtenido cdna que codifica el genoma di-a bajo el control del promotor inmediatamente temprano de citomegalovirus. Se ha localizado una secuencia viral situada en el gen de la replicasa que es t\u00f3xica en celulas procarioticas. Esta toxicidad se ha minimizado mediante el ensamblaje del cdna di-a como un cromosoma artificial de bacterias, que permite la amplificacion estable del cdna durante al menos 40 generaciones. la introducci\u00f3n en lar egion toxica de un intr\u00f3n de 133 nt, que es procesado en el 15-20% de las moleculas de rna en celulas st transfectadas, estabiliza el cdna durante al menos 120 generaciones.  3. Ensamblaje de un cdna que codifique un rna infectivo con el genoma completo del vgpt, y que genere un virus que se replique en los tractos enterico y respiratorio y sea virulento. Se ha clonado el genoma completo del vgpt con la seguncia del aislado pur-mad(atenuado, respiratorio y poco enterico), excepto el gen s, procedente del aislado pur-c11 (virulento, respiratorio y enterico),ya que datos previos indican que el gen s es un determinante del tropismo y la virulencia en el vgpt. Se han ensamblado las secuencias como un cromoso<\/p>\n<p>&nbsp;<\/p>\n<h3>Datos acad\u00e9micos de la tesis doctoral \u00ab<strong>Construccion de un cdna infectivo del coronavirus de la gastorenteritis porcina transmisible<\/strong>\u00ab<\/h3>\n<ul>\n<li><strong>T\u00edtulo de la tesis:<\/strong>\u00a0 Construccion de un cdna infectivo del coronavirus de la gastorenteritis porcina transmisible <\/li>\n<li><strong>Autor:<\/strong>\u00a0  Gonzalez Martienz Jos\u00e9 Manuel <\/li>\n<li><strong>Universidad:<\/strong>\u00a0 Aut\u00f3noma de Madrid<\/li>\n<li><strong>Fecha de lectura de la tesis:<\/strong>\u00a0 06\/10\/2000<\/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>Luis Enjanes Sanchez<\/li>\n<\/ul>\n<\/li>\n<li><strong>Tribunal<\/strong>\n<ul>\n<li>Presidente del tribunal: ricardo Flores pedauye <\/li>\n<li>Jos\u00e9 Antonio Melero fondevilla (vocal)<\/li>\n<li>esteban Domingo solans (vocal)<\/li>\n<li>agust\u00edn Portela moreira (vocal)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Tesis doctoral de Gonzalez Martienz Jos\u00e9 Manuel El objetivo final de este trabajo ha sido la obtenci\u00f3n de un cdna [&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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