{"id":90997,"date":"2018-03-11T10:09:36","date_gmt":"2018-03-11T10:09:36","guid":{"rendered":"https:\/\/www.deberes.net\/tesis\/sin-categoria\/molecular-characterization-of-cytosolic-alanine-aminotransferase-gene-expression-in-gilthead-sea-bream-sparus-aurata\/"},"modified":"2018-03-11T10:09:36","modified_gmt":"2018-03-11T10:09:36","slug":"molecular-characterization-of-cytosolic-alanine-aminotransferase-gene-expression-in-gilthead-sea-bream-sparus-aurata","status":"publish","type":"post","link":"https:\/\/www.deberes.net\/tesis\/bioquimica-molecular\/molecular-characterization-of-cytosolic-alanine-aminotransferase-gene-expression-in-gilthead-sea-bream-sparus-aurata\/","title":{"rendered":"Molecular characterization of cytosolic alanine aminotransferase gene expression in gilthead sea bream (sparus aurata)"},"content":{"rendered":"<h2>Tesis doctoral de <strong> Ida Guurtje Anemaet <\/strong><\/h2>\n<p>Molecular character1zation of cytosolic alanine aminotransferase gene expression in gilthead sea bream (sparus aurata) resumen: la alanina aminotransferasa (alt) cataliza de manera reversible la transferencia del grupo amino de la l-alanina al 2-oxoglutarato, dando lugar a la formaci\u00f3n de piruvato y l-glutamato. A fin de profundizar en el conocimiento de la funci\u00f3n ejercida por la isoforma citos\u00f3lica de la alt (calt) en dorada (sparus aurata), los objetivos de esta tesis han sido: i) clonar y caracterizar el promotor de calt de dorada; \u00c2\u00a1i) determinar los par\u00e1metros cin\u00e9ticos de las isoformas de calt; y iii) estudiar la regulaci\u00f3n nutricional y hormonal de la expresi\u00f3n de calt. El promotor de calt fue aislado mediante paseo cromos\u00f3mico sobre dna gen\u00f3mico de dorada. La caracterizaci\u00f3n de dicho promotor ha permitido delimitar el n\u00facleo m\u00ednimo con actividad promotora y el papel de diversos factores de transcripci\u00f3n y prote\u00ednas coactivadoras, como p300, c-myb y usf2a, sobre la actividad transcripcional del gen calt. Ensayos de rt-pcr y race pcr han permitido aislar una nueva isoenzima (calt2), que se genera por procesamiento alternativo del gen calt de dorada. La expresi\u00f3n de calt2 como una prote\u00edna fusionada a la prote\u00edna verde fluorescente (gfp) en c\u00e9lulas hek293 ha permitido demostrar, mediante microscop\u00eda confocal de fluorescencia, que calt2 localiza en el citosol. La funci\u00f3n metab\u00f3lica de las isoformas calt1 y calt2 en h\u00edgado de dorada se estudi\u00f3 mediante la caracterizaci\u00f3n cin\u00e9tica de ambas isoenzimas tras su expresi\u00f3n en saccharomyces cerevisiae, y el an\u00e1lisis de la modulaci\u00f3n de su expresi\u00f3n hep\u00e1tica en doradas sometidas a diversas condiciones nutricionales. El an\u00e1lisis cin\u00e9tico indica que calt2 es m\u00e1s eficaz en la cat\u00e1lisis de la conversi\u00f3n de l-alanina a piruvato que calt1. El ayuno estimula la expresi\u00f3n hep\u00e1tica de calt2, mientras que disminuye la de calt1. La alimentaci\u00f3n y el tratamiento con glucosa e insulina promueven los efectos contrarios al ayuno sobre la expresi\u00f3n de ambas isoenzimas. Considerados en su totalidad, los resultados obtenidos muestran que la expresi\u00f3n de calt2 aumenta en h\u00edgado en condiciones asociadas a gluconeog\u00e9nesis, mientras que la de calt1 est\u00e1 m\u00e1s relacionada con la utilizaci\u00f3n postprandial de los nutrientes de la dieta. Por otra parte, estudios recientes asocian un incremento de la actividad alt en sangre, dentro del rango considerado normal, con las fases iniciales del desarrollo de la diabetes de tipo 2 en humanos. El hecho de que nuestros estudios muestran un incremento de la expresi\u00f3n hep\u00e1tica de calt2 en h\u00edgado de doradas con diabetes inducida por tratamiento con estreptozotocina, sugiere que la expresi\u00f3n hep\u00e1tica de esta enzima podr\u00eda ser utilizada como un marcador eficaz en la patog\u00e9nesis de la diabetes en humanos.<\/p>\n<p>&nbsp;<\/p>\n<h3>Datos acad\u00e9micos de la tesis doctoral \u00ab<strong>Molecular characterization of cytosolic alanine aminotransferase gene expression in gilthead sea bream (sparus aurata)<\/strong>\u00ab<\/h3>\n<ul>\n<li><strong>T\u00edtulo de la tesis:<\/strong>\u00a0 Molecular characterization of cytosolic alanine aminotransferase gene expression in gilthead sea bream (sparus aurata) <\/li>\n<li><strong>Autor:<\/strong>\u00a0 Ida Guurtje Anemaet <\/li>\n<li><strong>Universidad:<\/strong>\u00a0 Barcelona<\/li>\n<li><strong>Fecha de lectura de la tesis:<\/strong>\u00a0 19\/12\/2008<\/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  Isabel V\u00e1zquez Baanante<\/li>\n<\/ul>\n<\/li>\n<li><strong>Tribunal<\/strong>\n<ul>\n<li>Presidente del tribunal: diego Haro bautista <\/li>\n<li>laura Baldoma llavines (vocal)<\/li>\n<li>Mar\u00eda Jos\u00e9 S\u00e1nchez-muros lozano (vocal)<\/li>\n<li>  (vocal)<\/li>\n<\/ul>\n<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Tesis doctoral de Ida Guurtje Anemaet Molecular character1zation of cytosolic alanine aminotransferase gene expression in gilthead sea bream (sparus aurata) 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