{"id":89786,"date":"2018-03-10T00:14:59","date_gmt":"2018-03-10T00:14:59","guid":{"rendered":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/pim1-una-map-quinasa-de-integridad-celular-en-pichia-pastoris-utilizacion-de-los-mutantes-pim1-para-la-liberacion-de-protea%c2%adnas-heterologas\/"},"modified":"2018-03-10T00:14:59","modified_gmt":"2018-03-10T00:14:59","slug":"pim1-una-map-quinasa-de-integridad-celular-en-pichia-pastoris-utilizacion-de-los-mutantes-pim1-para-la-liberacion-de-protea%c2%adnas-heterologas","status":"publish","type":"post","link":"https:\/\/www.deberes.net\/tesis\/ciencias-de-la-vida\/pim1-una-map-quinasa-de-integridad-celular-en-pichia-pastoris-utilizacion-de-los-mutantes-pim1-para-la-liberacion-de-protea%c2%adnas-heterologas\/","title":{"rendered":"Pim1, una map quinasa de integridad celular en pichia pastoris. utilizaci\u00f3n de los mutantes pim1 para la liberaci\u00f3n de prote\u00ednas heter\u00f3logas"},"content":{"rendered":"<h2>Tesis doctoral de <strong>  Cosano Maya Inmaculada C. <\/strong><\/h2>\n<p>La levadura metilotr\u00f3fica pichia pastoris ha sido empleada de manera satisfactoria para la expresi\u00f3n de prote\u00ednas heter\u00f3logas, tanto intracelulares como de secreci\u00f3n.Como sistema de expresi\u00f3n es muy similar a saccharomyces cerevisiae, pero presenta ciertas ventajas, entre ella sunas modificaciones psot-traduccionales m\u00e1s parecidas a las que tienen las c\u00e9lulas eucriotas superiores.  en nuestro laboratorio se ha desarrollado un sistema para la liberaci\u00f3n de prote\u00ednas heter\u00f3logas basado en el empleo de mutantes autol\u00edticos de s. Cerevisiae, remediables osmoticamente, que lisan a latas temperaturas. estos mutantes son deficientes en el gen slt2, una map quinasa responsable de la integridad celular.Dichos mutantes son capaces de liberar las prote\u00ednas intracelulares en ausencia de estabilizador osm\u00f3tico. Estas prote\u00ednas pueden ser separadas f\u00e1cilmente de los restos celulares por simple centrifugaci\u00f3n.  con el objeto de ampliar este sistema a p. Pastoris clonamos el gen hom\u00f3logo a slt2 (pim1) de esta levadura mediante hibridaci\u00f3n en colonia, empleando como sonda un fragmento de dicho gen amplificado por pcr, utilizando oligonuclotidos degenerados de zonas conservadas en map quinasas.  la prote\u00edna pim 1 de p. Pastoris presenta todas las caracter\u00edsticas estructurales de las prote\u00edna quinasas, as\u00ed como las secuencia caracter\u00edstica tey de regulaci\u00f3n de map quinasas. La interrupci\u00f3n de este gen origina el mismo fenotipo que el que presentna los mutantes slt2 de s. Cerevisiae; aut\u00f3lisis a 37\u00c2\u00bac y sensibilidad a caf\u00e9ina, ambos remediables en presencia de un estabilizador osm\u00f3tico. A pesar de que pim1 en p. Pastoris se activa en preencia de los mismo est\u00edmulos qu eactivan slt2 de s. Cerevisiae, el gen pim1 no complementa los defectos utantes slt 2 de cervisiae.  tras su incubaci\u00f3n a 37\u00c2\u00bac y un posterior choque osm\u00f3tico, los mutantes pim 1 que expresan la prote\u00edna modelo b-lactamasa liberan un 60% de la prote\u00edna heter<\/p>\n<p>&nbsp;<\/p>\n<h3>Datos acad\u00e9micos de la tesis doctoral \u00ab<strong>Pim1, una map quinasa de integridad celular en pichia pastoris. utilizaci\u00f3n de los mutantes pim1 para la liberaci\u00f3n de prote\u00ednas heter\u00f3logas<\/strong>\u00ab<\/h3>\n<ul>\n<li><strong>T\u00edtulo de la tesis:<\/strong>\u00a0 Pim1, una map quinasa de integridad celular en pichia pastoris. utilizaci\u00f3n de los mutantes pim1 para la liberaci\u00f3n de prote\u00ednas heter\u00f3logas <\/li>\n<li><strong>Autor:<\/strong>\u00a0  Cosano Maya Inmaculada C. <\/li>\n<li><strong>Universidad:<\/strong>\u00a0 Complutense de Madrid<\/li>\n<li><strong>Fecha de lectura de la tesis:<\/strong>\u00a0 17\/04\/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>Mar\u00eda Molina Mart\u00edn<\/li>\n<\/ul>\n<\/li>\n<li><strong>Tribunal<\/strong>\n<ul>\n<li>Presidente del tribunal: c\u00e9sar Nombela cano <\/li>\n<li> S\u00e1nchez-puelles gonz\u00e1lez-carvajal Jos\u00e9 Mar\u00eda (vocal)<\/li>\n<li>j. Heinisch jurgen (vocal)<\/li>\n<li>pablo Alvarez alvarez (vocal)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Tesis doctoral de Cosano Maya Inmaculada C. La levadura metilotr\u00f3fica pichia pastoris ha sido empleada de manera satisfactoria para la [&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 center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-gradient":""},"tablet":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-gradient":""},"mobile":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-gradient":""}},"ast-content-background-meta":{"desktop":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-gradient":""},"tablet":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-gradient":""},"mobile":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-gradient":""}},"footnotes":""},"categories":[35,278],"tags":[3243,186651,171388,24621,22663,186652],"class_list":["post-89786","post","type-post","status-publish","format-standard","hentry","category-ciencias-de-la-vida","category-microbiologia","tag-cesar-nombela-cano","tag-cosano-maya-inmaculada-c","tag-j-heinisch-jurgen","tag-maria-molina-martin","tag-pablo-alvarez-alvarez","tag-sanchez-puelles-gonzalez-carvajal-jose-maria"],"_links":{"self":[{"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/posts\/89786","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/comments?post=89786"}],"version-history":[{"count":0,"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/posts\/89786\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/media?parent=89786"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/categories?post=89786"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/tags?post=89786"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}