{"id":32085,"date":"2022-11-23T08:43:16","date_gmt":"2022-11-23T07:43:16","guid":{"rendered":"https:\/\/cab.inta-csic.es\/?post_type=tesis&#038;p=32085"},"modified":"2023-03-23T08:49:50","modified_gmt":"2023-03-23T07:49:50","slug":"32084-2","status":"publish","type":"tesis","link":"https:\/\/cab.inta-csic.es\/en\/tesis\/32084-2\/","title":{"rendered":"Exploraci\u00f3n de los mecanismos de adaptaci\u00f3n y resistencia a radiaci\u00f3n UV en microorganismos hal\u00f3filos extremos"},"content":{"rendered":"\n<p>El objetivo del estudio de esta tesis doctoral fue la exploraci\u00f3n de los mecanismos de resistencia y adaptaci\u00f3n a radiaci\u00f3n ultravioleta B (UVB) en microorganismos hal\u00f3filos extremos. Por una parte, para aislar genes implicados en resistencia a radiaci\u00f3n de estos microorganismos empleamos una t\u00e9cnica independiente de cultivo, la metagen\u00f3mica funcional. Para ello, se construyeron dos bibliotecas metagen\u00f3micas, empleando a Escherichia coli como hu\u00e9sped, con ADN (metagenoma) extra\u00eddo de salmueras de dos estanques cristalizadores de las salinas \u00bfBras del Port\u00bf (Santa Pola, Alicante), denominados CR30 y CCAB (39% y 30% de sal respectivamente). Mediante el cribado funcional de las bibliotecas metagen\u00f3micas se aislaron ocho clones resistentes a radiaci\u00f3n UVB. En los ocho fragmentos de ADN ambiental recuperados se identificaron quince genes de bacterias y arqueas responsables del fenotipo de resistencia a radiaci\u00f3n UVB. Estos genes codifican para prote\u00ednas anteriormente identificadas en la resistencia al da\u00f1o en el ADN, otras con funci\u00f3n conocida pero no relacionadas con resistencia a radiaci\u00f3n y varias prote\u00ednas con funci\u00f3n desconocida, similares a familias de prote\u00ednas hipot\u00e9ticas. Muchos de los genes tambi\u00e9n presentaron resistencia a 4-NQO, un compuesto que imita el da\u00f1o en el ADN que produce la radiaci\u00f3n UVB, por lo que esos genes podr\u00edan estar relacionados con protecci\u00f3n o reparaci\u00f3n del ADN; adem\u00e1s, algunos de ellos tambi\u00e9n presentaron resistencia al compuesto t\u00f3xico perclorato. Por otra parte, para comprender como responde y se adapta la maquinaria celular de hal\u00f3filos extremos a la radiaci\u00f3n UVB, se realiz\u00f3 un estudio transcript\u00f3mico de la bacteria Salinibacter ruber y la arquea Haloquadratum walsbyi, para analizar los cambios de expresi\u00f3n g\u00e9nica despu\u00e9s ser expuestos a una dosis subletal de radiaci\u00f3n UVB y recuperarse en presencia de luz u oscuridad para Sal. ruber y s\u00f3lo de luz para Hqr. walsbyi. En ambos microorganismos se observ\u00f3 la expresi\u00f3n diferencial de un porcentaje inferior al 20% de los genes\u2026<\/p>\n\n\n\n<div style=\"height:45px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<h5><strong>Exploring the mechanisms of adaptation and resistance to UV radiation in extreme halophilic microorganisms<\/strong><\/h5>\n\n\n\n<div style=\"height:20px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<p>The objective of this doctoral thesis was to explore the mechanisms of resistance and adaptation to ultraviolet B radiation (UVB) in extreme halophiles. On one hand, to isolate genes involved in radiation resistance from these microorganisms, we used a culture-independent technique, functional metagenomics. For this, two metagenomic libraries were constructed, using Escherichia coli as host, with DNA (metagenome) extracted from brines of two crystallizing ponds from &#8220;Bras del Port&#8221; saltern (Santa Pola, Alicante), called CR30 and CCAB (39% and 30% salt respectively). Eight UVB resistant clones were isolated by functional screening of metagenomic libraries. In the eight environmental DNA fragments recovered, fifteen bacteria and archaea genes responsible for the phenotype of UVB resistance were identified. These genes encode for proteins previously identified in DNA damage resistance, others with known function but not related to radiation resistance, and several proteins with unknown function, similar to families of hypothetical proteins. Many of the genes also showed resistance to 4-NQO, a compound that mimics the DNA damage produced by UVB radiation, so these genes could be related to DNA protection or repair; furthermore, some of them also showed resistance to the toxic compound perchlorate. On the other hand, to understand how the cellular machinery of extreme halophiles responds and adapts to UVB radiation, a transcriptomic study of the bacteria Salinibacter ruber and the archaea Haloquadratum walsbyi was carried out, in order to analyze the changes in gene expression after being exposed to a sublethal dose of UVB radiation and recovered in the presence of light or darkness for Sal. ruber, and only light for Hqr. walsbyi. In both microorganisms, the differential expression of less than 20% of the genes was demonstrated&#8230;.<\/p>\n","protected":false},"featured_media":0,"parent":0,"template":"","meta":[],"departamento_global":[493],"tesis_anos":[606],"acf":[],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v20.0 - https:\/\/yoast.com\/wordpress\/plugins\/seo\/ -->\n<title>Exploraci\u00f3n de los mecanismos de adaptaci\u00f3n y resistencia a radiaci\u00f3n UV en microorganismos hal\u00f3filos extremos &ndash; CAB<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/cab.inta-csic.es\/en\/tesis\/32084-2\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Exploraci\u00f3n de los mecanismos de adaptaci\u00f3n y resistencia a radiaci\u00f3n UV en microorganismos hal\u00f3filos extremos &ndash; CAB\" \/>\n<meta property=\"og:description\" content=\"El objetivo del estudio de esta tesis doctoral fue la exploraci\u00f3n de los mecanismos de resistencia y adaptaci\u00f3n a radiaci\u00f3n ultravioleta B (UVB) en microorganismos hal\u00f3filos extremos. 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