{"id":14599,"date":"2018-03-09T09:01:22","date_gmt":"2018-03-09T09:01:22","guid":{"rendered":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/purificacion-caracterizacion-y-clonacion-de-una-beta-16-glucanasa-de-trichoderma-harzianum\/"},"modified":"2018-03-09T09:01:22","modified_gmt":"2018-03-09T09:01:22","slug":"purificacion-caracterizacion-y-clonacion-de-una-beta-16-glucanasa-de-trichoderma-harzianum","status":"publish","type":"post","link":"https:\/\/www.deberes.net\/tesis\/ciencias-de-la-vida\/purificacion-caracterizacion-y-clonacion-de-una-beta-16-glucanasa-de-trichoderma-harzianum\/","title":{"rendered":"Purificaci\u00f3n, caracterizaci\u00f3n y clonaci\u00f3n de una beta-1,6-glucanasa de trichoderma harzianum"},"content":{"rendered":"<h2>Tesis doctoral de <strong> Manuel Montero Macarro <\/strong><\/h2>\n<p>Dentro del g\u00e9nero trichoderma se han encontrado numerosos agentes de control biol\u00f3gico frente a diversas enfermedades f\u00fangicas de inter\u00e9s en agricultura. Entre los mec\u00e1nimos por los que algunas cepas de trichoderma antagonizan a otros hongos filamentosos destaca el proceso del micoparasitismo, en el que juegan un importante papel las enzimas hidrol\u00edticas que trichoderma produce, entre las que se encuentran las beta-1,6-glucanasas. Hasta la fecha se han descrito dos isoenzimas con esta actividad en t.Harizanum.  en este trabajo hemos purificado y caracterizado una tercera isoenzima, a la que llamamos bgn16.3, que se ha purificado de cultivos de t.Harizanum cect 2413 con paredes celulares de botrytis cinerea como \u00fanica fuente de carbono.  la enzima fue purificada a homogeneidad electrofor\u00e9tica en tres pasos: afinidad, cromatoenfoque y filtraci\u00f3n en gel.  bgn16.3 present\u00f3 una masa molecular de 48 kda y un punto isoel\u00e9ctrico de 4.1 en cromatoenfoque y 4.5 en isoelectroenfoque. Adem\u00e1s se ha comprobado que posee efecto antil\u00fangico frente a penicillium digitatum tanto por s\u00ed sola como en combinaci\u00f3n con otras enzimas hidrol\u00edticas de trichoderma. a partir de secuencias proteicas parciales de bgn16.3, utilizando la t\u00e9cnica de pcr con oligonucle\u00f3tidos degenerados, se consigui\u00f3 clonar un fragmento del gen que la codifica.  utilizando este fragmento como sonda se realiz\u00f3 un escrutinio de una genoteca de cdna, consiguiendo un clon con el cdna completo de bgn16.3, que fue caracterizado por medios bioinform\u00e1ticos. Posteriormente tambi\u00e9n se clono el dna gen\u00f3mico de bgn16.3, incluyendo 1.6 kb de la regi\u00f3n promotora del gen. Se llevaron a cabo estudios de regulaci\u00f3n de la expresi\u00f3n del gen mediante northern-blot observando como car\u00e1cter\u00edsticas m\u00e1s interesantes la ausencia de expresi\u00f3n de la enzima en inducciones con quitina, sustrato habitualmente empleado para la obtenci\u00f3n de enzimas hidrol\u00edticas de trichoderma. mediant<\/p>\n<p>&nbsp;<\/p>\n<h3>Datos acad\u00e9micos de la tesis doctoral \u00ab<strong>Purificaci\u00f3n, caracterizaci\u00f3n y clonaci\u00f3n de una beta-1,6-glucanasa de trichoderma harzianum<\/strong>\u00ab<\/h3>\n<ul>\n<li><strong>T\u00edtulo de la tesis:<\/strong>\u00a0 Purificaci\u00f3n, caracterizaci\u00f3n y clonaci\u00f3n de una beta-1,6-glucanasa de trichoderma harzianum <\/li>\n<li><strong>Autor:<\/strong>\u00a0 Manuel Montero Macarro <\/li>\n<li><strong>Universidad:<\/strong>\u00a0 Salamanca<\/li>\n<li><strong>Fecha de lectura de la tesis:<\/strong>\u00a0 15\/12\/2001<\/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 Monte Vazquez<\/li>\n<\/ul>\n<\/li>\n<li><strong>Tribunal<\/strong>\n<ul>\n<li>Presidente del tribunal: andres Chordi corbo <\/li>\n<li>Santiago Gutierrez martin (vocal)<\/li>\n<li>Jos\u00e9 Mar\u00eda Medina jimenez (vocal)<\/li>\n<li>matteo Lorito (vocal)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Tesis doctoral de Manuel Montero Macarro Dentro del g\u00e9nero trichoderma se han encontrado numerosos agentes de control biol\u00f3gico frente a [&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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