{"id":13164,"date":"2001-05-10T00:00:00","date_gmt":"2001-05-10T00:00:00","guid":{"rendered":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/localizacion-separacion-e-identificacion-del-complejo-nadph-deshidrogenasa-de-los-cloroplastos-de-cebadahordeum-vulgare-l\/"},"modified":"2001-05-10T00:00:00","modified_gmt":"2001-05-10T00:00:00","slug":"localizacion-separacion-e-identificacion-del-complejo-nadph-deshidrogenasa-de-los-cloroplastos-de-cebadahordeum-vulgare-l","status":"publish","type":"post","link":"https:\/\/www.deberes.net\/tesis\/ciencias-de-la-vida\/localizacion-separacion-e-identificacion-del-complejo-nadph-deshidrogenasa-de-los-cloroplastos-de-cebadahordeum-vulgare-l\/","title":{"rendered":"Localizacion, separacion e identificacion del complejo nad(p)h deshidrogenasa de los cloroplastos de cebada(hordeum vulgare l.)"},"content":{"rendered":"<h2>Tesis doctoral de <strong> Araceli Garcia Buendia <\/strong><\/h2>\n<p>Con el proposito de localizar, aislar e identificar el complejo nad(p)h deshidrogenasa (nad(p)h dh) de los cloroplastos de cebada, se parti\u00f3 de tilacoides estromales obtenidos por dos metodos de fraccionamiento. Uno basado en la solubilizaci\u00f3n de las membranas tilacoidales con el detergente digitonina y el otro en el tratamiento fisico de las mismas con ultrasonidos o sonicaci\u00f3n.  el estudio de las fracciones obtenidas, mediante parametros quimicos y bioquimicos,nos permiti\u00f3 comprobar la pureza de las muestras asi como la localizacion preferente en los tilacoides estromales del complejo objeto de estudio.  para aislar el complejo a partir de la muestra de tilacoides estromales, utilizamos la electroforesis nativa con triton x-100 y posterior tinci\u00f3n con nadh y nitroazul de tetrazolio(nadh-nbt), obteniendo tres bandas con actividad nadh-nbtr(bandas 1,2 y 3). La obtenci\u00f3n de tres bandas nos llev\u00f3 a pensar que el complejo se disociaba durante la electroforesis. A continuaci\u00f3n las bandas se electroeluyeron con el fin de obtener los perfiles polipeptidicos para su analisis por inmunotransferencia. El an\u00e1lisis con el anticuerpo anti-ndha permiti\u00f3 identificar la enzima electroeluida de la banda 2, como el complejo nadh dh tilacoidal. Tambian se comprob\u00f3 la asociaci\u00f3n del complejo a la enzima fnr utilizando el anticuerpo fabricado contra fnr de cebada.  para intentar resolver el problema de la disociaci\u00f3n del complejo, utilizamos para su aislamiento la electroforesis nativa con azul coomassie(bn-page) partiendo de tilacoides estromales obtenidos con sonicaci\u00f3n. Despues de la electroforesis y tinci\u00f3n con nad(p)h-nbt, se obtuvo una sola banda con actividad que se analizo por inmunotransferencia con los anticuerpos anti-ndha,anti-ndhh y anti-polipeptido de 51 kda, identificandola como el complejo nad(p)h dh cloroplastico. Antes descartamos posibles contaminaciones por el complejo atp sintetasa y la enzima rubisco.  una vez aislado<\/p>\n<p>&nbsp;<\/p>\n<h3>Datos acad\u00e9micos de la tesis doctoral \u00ab<strong>Localizacion, separacion e identificacion del complejo nad(p)h deshidrogenasa de los cloroplastos de cebada(hordeum vulgare l.)<\/strong>\u00ab<\/h3>\n<ul>\n<li><strong>T\u00edtulo de la tesis:<\/strong>\u00a0 Localizacion, separacion e identificacion del complejo nad(p)h deshidrogenasa de los cloroplastos de cebada(hordeum vulgare l.) <\/li>\n<li><strong>Autor:<\/strong>\u00a0 Araceli Garcia Buendia <\/li>\n<li><strong>Universidad:<\/strong>\u00a0 Murcia<\/li>\n<li><strong>Fecha de lectura de la tesis:<\/strong>\u00a0 05\/10\/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>Juan Cuello Moreno<\/li>\n<\/ul>\n<\/li>\n<li><strong>Tribunal<\/strong>\n<ul>\n<li>Presidente del tribunal: romualdo Mu\u00f1oz giron <\/li>\n<li>Jos\u00e9 Egea caballero (vocal)<\/li>\n<li>francisca Sevilla valenzuela (vocal)<\/li>\n<li>ramon Madrid vicente (vocal)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Tesis doctoral de Araceli Garcia Buendia Con el proposito de localizar, aislar e identificar el complejo nad(p)h deshidrogenasa (nad(p)h dh) [&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":[972,35,479,8235],"tags":[42832,8466,25006,8432,8433,8397],"class_list":["post-13164","post","type-post","status-publish","format-standard","hentry","category-botanica","category-ciencias-de-la-vida","category-fisiologia-vegetal","category-murcia","tag-araceli-garcia-buendia","tag-francisca-sevilla-valenzuela","tag-jose-egea-caballero","tag-juan-cuello-moreno","tag-ramon-madrid-vicente","tag-romualdo-munoz-giron"],"_links":{"self":[{"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/posts\/13164","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=13164"}],"version-history":[{"count":0,"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/posts\/13164\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/media?parent=13164"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/categories?post=13164"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.deberes.net\/tesis\/wp-json\/wp\/v2\/tags?post=13164"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}