{"id":2445,"date":"2023-03-15T08:37:59","date_gmt":"2023-03-15T14:37:59","guid":{"rendered":"https:\/\/nuevo22.cidsamexico.com\/?page_id=2445"},"modified":"2023-04-20T12:42:05","modified_gmt":"2023-04-20T17:42:05","slug":"gst-tag-definicion-y-datos","status":"publish","type":"page","link":"https:\/\/nuevo22.cidsamexico.com\/index.php\/gst-tag-definicion-y-datos\/","title":{"rendered":"GST-Tag : Definici\u00f3n y Datos"},"content":{"rendered":"\t\t<div data-elementor-type=\"wp-post\" data-elementor-id=\"2445\" class=\"elementor elementor-2445\">\n\t\t\t\t\t\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-759c9a9 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"759c9a9\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-a00c95f\" data-id=\"a00c95f\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-081e1e5 elementor-widget elementor-widget-text-editor\" data-id=\"081e1e5\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<style>\/*! elementor - v3.7.8 - 02-10-2022 *\/\n.elementor-widget-text-editor.elementor-drop-cap-view-stacked .elementor-drop-cap{background-color:#818a91;color:#fff}.elementor-widget-text-editor.elementor-drop-cap-view-framed .elementor-drop-cap{color:#818a91;border:3px solid;background-color:transparent}.elementor-widget-text-editor:not(.elementor-drop-cap-view-default) .elementor-drop-cap{margin-top:8px}.elementor-widget-text-editor:not(.elementor-drop-cap-view-default) .elementor-drop-cap-letter{width:1em;height:1em}.elementor-widget-text-editor .elementor-drop-cap{float:left;text-align:center;line-height:1;font-size:50px}.elementor-widget-text-editor .elementor-drop-cap-letter{display:inline-block}<\/style>\t\t\t\t<p>Como una proteina ayuda a purificar a otra proteina\u00a0<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-7cfca4c elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"7cfca4c\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-d5c7869\" data-id=\"d5c7869\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-a1ae3e5 elementor-widget elementor-widget-image\" data-id=\"a1ae3e5\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<style>\/*! elementor - v3.7.8 - 02-10-2022 *\/\n.elementor-widget-image{text-align:center}.elementor-widget-image a{display:inline-block}.elementor-widget-image a img[src$=\".svg\"]{width:48px}.elementor-widget-image img{vertical-align:middle;display:inline-block}<\/style>\t\t\t\t\t\t\t\t\t<figure class=\"wp-caption\">\n\t\t\t\t\t\t\t\t\t\t<img width=\"718\" height=\"345\" src=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag2.jpg\" class=\"attachment-large size-large\" alt=\"\" loading=\"lazy\" srcset=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag2.jpg 718w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag2-300x144.jpg 300w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag2-600x288.jpg 600w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag2-280x135.jpg 280w\" sizes=\"(max-width: 718px) 100vw, 718px\" \/>\t\t\t\t\t\t\t\t\t\t\t<figcaption class=\"widget-image-caption wp-caption-text\">Figura 1: La etiqueta GST y sus caracteristicas<\/figcaption>\n\t\t\t\t\t\t\t\t\t\t<\/figure>\n\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-3e2caac elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"3e2caac\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-ea4d956\" data-id=\"ea4d956\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-29c62a7 elementor-widget elementor-widget-spacer\" data-id=\"29c62a7\" data-element_type=\"widget\" data-widget_type=\"spacer.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<style>\/*! elementor - v3.7.8 - 02-10-2022 *\/\n.elementor-column .elementor-spacer-inner{height:var(--spacer-size)}.e-container{--container-widget-width:100%}.e-container>.elementor-widget-spacer{width:var(--container-widget-width,var(--spacer-size));-ms-flex-item-align:stretch;align-self:stretch;-ms-flex-negative:0;flex-shrink:0}.e-container>.elementor-widget-spacer>.elementor-widget-container,.e-container>.elementor-widget-spacer>.elementor-widget-container>.elementor-spacer{height:100%}.e-container>.elementor-widget-spacer>.elementor-widget-container>.elementor-spacer>.elementor-spacer-inner{height:var(--container-widget-height,var(--spacer-size))}<\/style>\t\t<div class=\"elementor-spacer\">\n\t\t\t<div class=\"elementor-spacer-inner\"><\/div>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-ffb93e0 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"ffb93e0\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-14bd3a7\" data-id=\"14bd3a7\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-063ec90 elementor-widget elementor-widget-text-editor\" data-id=\"063ec90\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>\u00bfQu\u00e9 es la etiqueta GST?<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-4d34c2d elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"4d34c2d\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-771537b\" data-id=\"771537b\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-ec61c5b elementor-widget elementor-widget-text-editor\" data-id=\"ec61c5b\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Existen m\u00faltiples etiquetas de afinidad de prote\u00ednas que se utilizan para la purificaci\u00f3n de prote\u00ednas y la etiqueta GST es una de ellas. GST es la abreviatura de glutati\u00f3n-S-transferasa y se refiere a una prote\u00edna completa, no solo a unos pocos amino\u00e1cidos como la etiqueta Rho1D4. La etiqueta de la enzima GST se ha utilizado para la purificaci\u00f3n de prote\u00ednas desde finales de la d\u00e9cada de 1980 (Smith &amp; Johnson 1988) y se ha establecido como una forma confiable para los ensayos de purificaci\u00f3n de prote\u00ednas que requieren el m\u00e1s alto nivel de pureza (Harper &amp; Speicher 2013).<\/p><p><strong>El tama\u00f1o de la etiqueta GST es de 26 kDa<\/strong> y, por lo tanto, es realmente grande en comparaci\u00f3n con otras etiquetas de afinidad de prote\u00ednas. En teor\u00eda, se puede fusionar con el extremo C o N de una prote\u00edna (Terpe 2003), pero <strong>recomendamos enf\u00e1ticamente<\/strong> agregarlo al extremo N. Hasta ahora se ha utilizado con \u00e9xito para purificar prote\u00ednas de c\u00e9lulas bacterianas (Smith y Johnson 1988), levaduras (Lu et al. 1997) y c\u00e9lulas de mam\u00edferos (Rudert et al. 1996). Sus caracter\u00edsticas generales y secuencia se pueden encontrar aqu\u00ed.<\/p><p>Ahora sabemos que la etiqueta GST b\u00e1sicamente es una prote\u00edna completa que se une a la prote\u00edna de inter\u00e9s. Pero ahora, \u00bfqu\u00e9 es exactamente la glutati\u00f3n-S-transferasa?<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-abc8a9b elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"abc8a9b\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-f310a18\" data-id=\"f310a18\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-86ef726 elementor-widget-divider--view-line elementor-widget elementor-widget-divider\" data-id=\"86ef726\" data-element_type=\"widget\" data-widget_type=\"divider.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t<style>\/*! elementor - v3.7.8 - 02-10-2022 *\/\n.elementor-widget-divider{--divider-border-style:none;--divider-border-width:1px;--divider-color:#2c2c2c;--divider-icon-size:20px;--divider-element-spacing:10px;--divider-pattern-height:24px;--divider-pattern-size:20px;--divider-pattern-url:none;--divider-pattern-repeat:repeat-x}.elementor-widget-divider .elementor-divider{display:-webkit-box;display:-ms-flexbox;display:flex}.elementor-widget-divider .elementor-divider__text{font-size:15px;line-height:1;max-width:95%}.elementor-widget-divider .elementor-divider__element{margin:0 var(--divider-element-spacing);-ms-flex-negative:0;flex-shrink:0}.elementor-widget-divider .elementor-icon{font-size:var(--divider-icon-size)}.elementor-widget-divider .elementor-divider-separator{display:-webkit-box;display:-ms-flexbox;display:flex;margin:0;direction:ltr}.elementor-widget-divider--view-line_icon .elementor-divider-separator,.elementor-widget-divider--view-line_text .elementor-divider-separator{-webkit-box-align:center;-ms-flex-align:center;align-items:center}.elementor-widget-divider--view-line_icon .elementor-divider-separator:after,.elementor-widget-divider--view-line_icon .elementor-divider-separator:before,.elementor-widget-divider--view-line_text .elementor-divider-separator:after,.elementor-widget-divider--view-line_text .elementor-divider-separator:before{display:block;content:\"\";border-bottom:0;-webkit-box-flex:1;-ms-flex-positive:1;flex-grow:1;border-top:var(--divider-border-width) var(--divider-border-style) var(--divider-color)}.elementor-widget-divider--element-align-left .elementor-divider .elementor-divider-separator>.elementor-divider__svg:first-of-type{-webkit-box-flex:0;-ms-flex-positive:0;flex-grow:0;-ms-flex-negative:100;flex-shrink:100}.elementor-widget-divider--element-align-left .elementor-divider-separator:before{content:none}.elementor-widget-divider--element-align-left .elementor-divider__element{margin-left:0}.elementor-widget-divider--element-align-right .elementor-divider .elementor-divider-separator>.elementor-divider__svg:last-of-type{-webkit-box-flex:0;-ms-flex-positive:0;flex-grow:0;-ms-flex-negative:100;flex-shrink:100}.elementor-widget-divider--element-align-right .elementor-divider-separator:after{content:none}.elementor-widget-divider--element-align-right .elementor-divider__element{margin-right:0}.elementor-widget-divider:not(.elementor-widget-divider--view-line_text):not(.elementor-widget-divider--view-line_icon) .elementor-divider-separator{border-top:var(--divider-border-width) var(--divider-border-style) var(--divider-color)}.elementor-widget-divider--separator-type-pattern{--divider-border-style:none}.elementor-widget-divider--separator-type-pattern.elementor-widget-divider--view-line .elementor-divider-separator,.elementor-widget-divider--separator-type-pattern:not(.elementor-widget-divider--view-line) .elementor-divider-separator:after,.elementor-widget-divider--separator-type-pattern:not(.elementor-widget-divider--view-line) .elementor-divider-separator:before,.elementor-widget-divider--separator-type-pattern:not([class*=elementor-widget-divider--view]) .elementor-divider-separator{width:100%;min-height:var(--divider-pattern-height);-webkit-mask-size:var(--divider-pattern-size) 100%;mask-size:var(--divider-pattern-size) 100%;-webkit-mask-repeat:var(--divider-pattern-repeat);mask-repeat:var(--divider-pattern-repeat);background-color:var(--divider-color);-webkit-mask-image:var(--divider-pattern-url);mask-image:var(--divider-pattern-url)}.elementor-widget-divider--no-spacing{--divider-pattern-size:auto}.elementor-widget-divider--bg-round{--divider-pattern-repeat:round}.rtl .elementor-widget-divider .elementor-divider__text{direction:rtl}.e-container>.elementor-widget-divider{width:var(--container-widget-width,100%);--flex-grow:var(--container-widget-flex-grow,0)}<\/style>\t\t<div class=\"elementor-divider\">\n\t\t\t<span class=\"elementor-divider-separator\">\n\t\t\t\t\t\t<\/span>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-df28bd5 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"df28bd5\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-2b45040\" data-id=\"2b45040\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-675de9c elementor-widget elementor-widget-text-editor\" data-id=\"675de9c\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>\u00bfQu\u00e9 es GST \/ Glutati\u00f3n-S-Transferasa?<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-c14c44f elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"c14c44f\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-53e3006\" data-id=\"53e3006\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-0b68545 elementor-widget elementor-widget-text-editor\" data-id=\"0b68545\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Las glutati\u00f3n-S-transferasas son una familia de enzimas procariotas y eucariotas que participan principalmente en la desintoxicaci\u00f3n de las c\u00e9lulas y el cuerpo. En los casos m\u00e1s extremos, las prote\u00ednas GST pueden constituir hasta el 10 % del proteoma citos\u00f3lico completo de algunas c\u00e9lulas de mam\u00edferos (Oakley 2011). La reacci\u00f3n que catalizan las glutati\u00f3n S-transferasas es la oxidaci\u00f3n del glutati\u00f3n reducido, tambi\u00e9n llamado GSH. La funci\u00f3n principal de GST en la c\u00e9lula es la protecci\u00f3n del ADN y las prote\u00ednas celulares de sustancias t\u00f3xicas externas. El mecanismo cataliza un ataque nucleof\u00edlico sobre la sustancia t\u00f3xica. Al activar un grupo tiol de GSH, reacciona con grupos adecuados sobre las sustancias t\u00f3xicas. Este acoplamiento de GSH a las toxinas conduce a su excreci\u00f3n de la c\u00e9lula oa una mayor procesi\u00f3n metab\u00f3lica (Hayes et al. 2004). La alta especificidad de GST hacia el glutati\u00f3n reducido (GSH) es la raz\u00f3n de la alta pureza de prote\u00ednas de los ensayos de purificaci\u00f3n.<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-3018743 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"3018743\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-e9535b0\" data-id=\"e9535b0\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-2df3494 elementor-widget elementor-widget-image\" data-id=\"2df3494\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t<figure class=\"wp-caption\">\n\t\t\t\t\t\t\t\t\t\t<img width=\"800\" height=\"566\" src=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-Function2.jpg\" class=\"attachment-large size-large\" alt=\"\" loading=\"lazy\" srcset=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-Function2.jpg 1015w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-Function2-300x212.jpg 300w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-Function2-768x543.jpg 768w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-Function2-600x424.jpg 600w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-Function2-226x160.jpg 226w\" sizes=\"(max-width: 800px) 100vw, 800px\" \/>\t\t\t\t\t\t\t\t\t\t\t<figcaption class=\"widget-image-caption wp-caption-text\">Fig. 2: Proceso de desintoxicaci\u00f3n por Glutati\u00f3n-S-Transferasas<\/figcaption>\n\t\t\t\t\t\t\t\t\t\t<\/figure>\n\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-bf50614 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"bf50614\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-ad60ab8\" data-id=\"ad60ab8\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-020b1d2 elementor-widget elementor-widget-text-editor\" data-id=\"020b1d2\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Como se indic\u00f3: existen numerosos ort\u00f3logos de GST, pero se eligi\u00f3 uno espec\u00edfico como plantilla para la etiqueta GST. Entonces, \u00bfpor qu\u00e9 GST de Schistosoma japonicum?<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-3ece7db elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"3ece7db\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-adc052e\" data-id=\"adc052e\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-51445f1 elementor-widget-divider--view-line elementor-widget elementor-widget-divider\" data-id=\"51445f1\" data-element_type=\"widget\" data-widget_type=\"divider.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-divider\">\n\t\t\t<span class=\"elementor-divider-separator\">\n\t\t\t\t\t\t<\/span>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-6e0748e elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"6e0748e\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-fd2a999\" data-id=\"fd2a999\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<section class=\"elementor-section elementor-inner-section elementor-element elementor-element-c452cb7 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"c452cb7\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-50 elementor-inner-column elementor-element elementor-element-e1b9ec7\" data-id=\"e1b9ec7\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-b78cda0 elementor-widget elementor-widget-text-editor\" data-id=\"b78cda0\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>\u00bfPor qu\u00e9 GST de Schistosoma japonicum?<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-e508e33 elementor-widget elementor-widget-text-editor\" data-id=\"e508e33\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Las glutati\u00f3n-S-transferasas se pueden encontrar en organismos que van desde procariotas hasta eucariotas (Allocati et al. 2008). Entonces, \u00bfpor qu\u00e9 se usa esta prote\u00edna GST espec\u00edfica para la purificaci\u00f3n por afinidad? \u00bfCu\u00e1les son sus ventajas? Esta respuesta se remonta a la primera descripci\u00f3n de GST como una opci\u00f3n para la cromatograf\u00eda de afinidad y la purificaci\u00f3n de prote\u00ednas. Como se cita formalmente, la primera menci\u00f3n de GST para la purificaci\u00f3n de prote\u00ednas fue en 1988 por Smith &amp; Johnson. Antes de eso Smith et al. publicaron un art\u00edculo en el que describ\u00edan el posible uso de GST como vacuna contra Schistosoma japonicum.<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t<div class=\"elementor-column elementor-col-50 elementor-inner-column elementor-element elementor-element-513962c\" data-id=\"513962c\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-b8add2b elementor-widget elementor-widget-image\" data-id=\"b8add2b\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t<figure class=\"wp-caption\">\n\t\t\t\t\t\t\t\t\t\t<img width=\"353\" height=\"408\" src=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/Sketch-S.jpg\" class=\"attachment-large size-large\" alt=\"\" loading=\"lazy\" srcset=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/Sketch-S.jpg 353w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/Sketch-S-260x300.jpg 260w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/Sketch-S-138x160.jpg 138w\" sizes=\"(max-width: 353px) 100vw, 353px\" \/>\t\t\t\t\t\t\t\t\t\t\t<figcaption class=\"widget-image-caption wp-caption-text\">Fig. 3: Imagen de Schistosoma japonicum<\/figcaption>\n\t\t\t\t\t\t\t\t\t\t<\/figure>\n\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-d46bbfe elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"d46bbfe\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-8646944\" data-id=\"8646944\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-b137a79 elementor-widget elementor-widget-text-editor\" data-id=\"b137a79\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Schistosoma japonicum, un Platyhelminthes, es una de las principales causas de la enfermedad esquistosomiasis y la prote\u00edna GST de dicha especie promet\u00eda ser la base para una vacuna contra Schistosoma japonicum (Smith et al. 1988). Debido a este estudio, Smith ya trabaj\u00f3 con este ort\u00f3logo espec\u00edfico de la prote\u00edna GST y, por lo tanto, sigui\u00f3 siendo la base para el m\u00e9todo de purificaci\u00f3n de prote\u00ednas descrito m\u00e1s tarde. Y desde 1988, GST de Schistosoma japonicum permaneci\u00f3 como el ort\u00f3logo de GST que se usa como etiqueta de afinidad de GST.<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-831c35e elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"831c35e\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-6d4525f\" data-id=\"6d4525f\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-26e3a58 elementor-widget-divider--view-line elementor-widget elementor-widget-divider\" data-id=\"26e3a58\" data-element_type=\"widget\" data-widget_type=\"divider.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-divider\">\n\t\t\t<span class=\"elementor-divider-separator\">\n\t\t\t\t\t\t<\/span>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-bdc6b8f elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"bdc6b8f\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-cf39799\" data-id=\"cf39799\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-ce511ea elementor-widget elementor-widget-text-editor\" data-id=\"ce511ea\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Caracter\u00edsticas de GST-Tag<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-58b5e54 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"58b5e54\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-0ce2c16\" data-id=\"0ce2c16\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-9dd863b elementor-widget elementor-widget-text-editor\" data-id=\"9dd863b\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Recopilamos los datos m\u00e1s importantes sobre la etiqueta GST y sus capacidades para la purificaci\u00f3n de prote\u00ednas.<br \/><strong>Nota:<\/strong> enumeramos la secuencia &#8220;est\u00e1ndar&#8221; utilizada en los vectores pGEX.<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-1d2a2f9 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"1d2a2f9\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-b37261a\" data-id=\"b37261a\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-73a1e53 elementor-widget elementor-widget-text-editor\" data-id=\"73a1e53\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Tabla 1: Resumen de las caracter\u00edsticas m\u00e1s importantes de la etiqueta GST<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-8c94139 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"8c94139\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-1846ed3\" data-id=\"1846ed3\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-3679504 elementor-widget elementor-widget-text-editor\" data-id=\"3679504\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<table class=\"col-xl-9 col-sm-12 responsiv py-auto mb-3\"><tbody><tr><th colspan=\"2\">FEATURES OF THE GST-TAG:<\/th><\/tr><tr><td>Amino acid Sequence<\/td><td>MSPILGYWKIKGLVQPTRLLLEYLEEKYEE<br \/>HLYERDEGDKWRNKKFELGLEFPNLPYYID<br \/>GDVKLTQSMAIIRYIADKHNMLGGCPKERA<br \/>EISMLEGAVLDIRYGVSRIAYSKDFETLKV<br \/>DFLSKLPEMLKMFEDRLCHKTYLNGDHVTH<br \/>PDFMLYDALDVVLYMDPMCLDAFPKLVCFK<br \/>KRIEAIPQIDKYLKSSKYIAWPLQGWQATF<br \/>GGGDHPPKSD<\/td><\/tr><tr><td>DNA sequence<br \/>(Based on pGEX-4T1 vector)<\/td><td>ATG TCCCCTATAC TAGGTTATTG GAAAATTAAG<br \/>GGCCTTGTGC AACCCACTCG ACTTCTTTTG<br \/>GAATATCTTG AAGAAAAATA TGAAGAGCAT<br \/>TTGTATGAGC GCGATGAAGG TGATAAATGG<br \/>CGAAACAAAA AGTTTGAATT GGGTTTGGAG<br \/>TTTCCCAATC TTCCTTATTA TATTGATGGT<br \/>GATGTTAAAT TAACACAGTC TATGGCCATC<br \/>ATACGTTATA TAGCTGACAA GCACAACATG<br \/>TTGGGTGGTT GTCCAAAAGA GCGTGCAGAG<br \/>ATTTCAATGC TTGAAGGAGC GGTTTTGGAT<br \/>ATTAGATACG GTGTTTCGAG AATTGCATAT<br \/>AGTAAAGACT TTGAAACTCT CAAAGTTGAT<br \/>TTTCTTAGCA AGCTACCTGA AATGCTGAAA<br \/>ATGTTCGAAG ATCGTTTATG TCATAAAACA<br \/>TATTTAAATG GTGATCATGT AACCCATCCT<br \/>GACTTCATGT TGTATGACGC TCTTGATGTT<br \/>GTTTTATACA TGGACCCAAT GTGCCTGGAT<br \/>GCGTTCCCAA AATTAGTTTG TTTTAAAAAA<br \/>CGTATTGAAG CTATCCCACA AATTGATAAG<br \/>TACTTGAAAT CCAGCAAGTA TATAGCATGG<br \/>CCTTTGCAGG GCTGGCAAGC CACGTTTGGT<br \/>GGTGGCGACC ATCCTCCAAA ATCGGATCTG<br \/>GTTCCGCGTG GATCCCCGGA ATTCCCGGGT<br \/>CGACTCGAGC GGCCGCATCG TGACTGA<\/td><\/tr><tr><td>Size<\/td><td>26 kDa<\/td><\/tr><tr><td>Compatibility to recombinant proteins<\/td><td>N-terminus is highly recommended<\/td><\/tr><tr><td>Combinable with other affinity tags?<\/td><td>Yes, see our\u00a0<a href=\"https:\/\/cube-biotech.com\/products\/active-proteins\/soluble-proteins\/gst-egfp-fusion-protein-rho1d4-tag\/29911\">GST-and-Rho-tagged eGFP<\/a>\u00a0as an example<\/td><\/tr><tr><td>Specificity of interaction (KD)<\/td><td>1 \u00b5M<\/td><\/tr><tr><td>Typical protein yield per ml of high-quality purification resin.<\/td><td>10-12 mg\/ml<\/td><\/tr><tr><td>Elution conditions<\/td><td><a href=\"https:\/\/cube-biotech.com\/products\/protein-purification-products\/gst-tag-affinity-resins-magbeads\/l-glutathione-reduced\/61035\">Reduced Glutathione<\/a>, pH shift, or tag cleavage<\/td><\/tr><tr><td>Binds to purification resin \/ magnetic bead via<\/td><td><a href=\"https:\/\/cube-biotech.com\/products\/protein-purification-products\/gst-tag-affinity-resins-magbeads\/l-glutathione-reduced\/61035\">Reduced Glutathione<\/a><\/td><\/tr><tr><td>Suitable protein purification methods<\/td><td><a href=\"https:\/\/cube-biotech.com\/products\/protein-purification-products\/gst-tag-affinity-resins-magbeads\/\">Agarose resin &amp; magnetic beads<\/a><\/td><\/tr><\/tbody><\/table>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-a8e1b11 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"a8e1b11\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-cb3d94e\" data-id=\"cb3d94e\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-c9bbbe2 elementor-widget-divider--view-line elementor-widget elementor-widget-divider\" data-id=\"c9bbbe2\" data-element_type=\"widget\" data-widget_type=\"divider.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-divider\">\n\t\t\t<span class=\"elementor-divider-separator\">\n\t\t\t\t\t\t<\/span>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-cb3713b elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"cb3713b\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-c88aad2\" data-id=\"c88aad2\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-7375ac2 elementor-widget elementor-widget-text-editor\" data-id=\"7375ac2\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>C\u00f3mo agregar\/clonar una etiqueta GST en un pl\u00e1smido<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-d50ebac elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"d50ebac\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-486142f\" data-id=\"486142f\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<section class=\"elementor-section elementor-inner-section elementor-element elementor-element-ead00a4 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"ead00a4\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-50 elementor-inner-column elementor-element elementor-element-5ccd466\" data-id=\"5ccd466\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-f90d28f elementor-widget elementor-widget-text-editor\" data-id=\"f90d28f\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>La mejor manera de agregar una etiqueta GST a una prote\u00edna de inter\u00e9s es usar un vector de la serie pGEX. Tambi\u00e9n vienen con diferentes sitios de escisi\u00f3n de prote\u00ednas. Estos son importantes para los m\u00e9todos mencionados m\u00e1s adelante para eliminar una etiqueta GST. La Figura 4 muestra el proceso y los pasos de nuestra estrategia de clonaci\u00f3n recomendada.<\/p><p><strong>1.<\/strong> Todos los vectores pGEX vienen con un sitio de clonaci\u00f3n m\u00faltiple (MSC) que contiene m\u00faltiples sitios de digesti\u00f3n con enzimas de restricci\u00f3n. Estos var\u00edan de vector pGEX a vector. La Tabla 2 proporciona una descripci\u00f3n general aqu\u00ed. Adem\u00e1s, todos los vectores pGEX contienen tres codones de parada que cubren los tres marcos de lectura posibles en una cepa de ADN, despu\u00e9s de la MSC. Esto es para asegurar que la prote\u00edna expresada no tenga que contener el cod\u00f3n de terminaci\u00f3n por s\u00ed misma.<\/p><p><strong>2.<\/strong> El siguiente paso es crear un producto de PCR de su gen\/prote\u00edna de inter\u00e9s que est\u00e9 flanqueado en ambas direcciones con sitios de restricci\u00f3n que est\u00e1n presentes en el MCS del vector pGEX elegido. Estos manuales deben cubrir cada uno:<br \/>\u00a0 \u00a0 1. Voladizo de 3-4 nts (cuanto m\u00e1s largo, mejor para la digesti\u00f3n de restricci\u00f3n, pero sugerimos 3-4)<br \/>\u00a0 \u00a0 2. El sitio de restricci\u00f3n<br \/>\u00a0 \u00a0 3. 20-25 nts de tu gen de inter\u00e9s<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-ed83d49 elementor-widget elementor-widget-text-editor\" data-id=\"ed83d49\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p><strong>Perm\u00edtanos darle un ejemplo de esto: intentemos asumir que desea agregar una etiqueta GST a eGFP.<\/strong><\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-3d0d7d8 elementor-widget elementor-widget-text-editor\" data-id=\"3d0d7d8\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<div class=\"col-6\"><p>\u00a0<\/p><p>Legend &#8211; Forward primer:<br \/><strong>Overhang<\/strong><br \/><span style=\"color: #008000;\">Restriction site -&gt; BamHI in this case<\/span><br \/><span style=\"color: #ff6600;\">First 20 nts of eGFP<\/span><\/p><p>5&#8242;-<strong>GGG<\/strong>\u00a0<span style=\"color: #008000;\">GGATCC<\/span>\u00a0<span style=\"color: #ff6600;\">ATGAAACATCACCATCACCA<\/span>-3&#8242;<\/p><\/div><div>\u00a0<\/div><div>\u00a0<\/div><div class=\"col-6\"><p>Legend &#8211; reverse Primer:<br \/>Overhang<br \/><span style=\"color: #008000;\">Restriction site -&gt; EcoRI in this case, reverse complement<\/span><br \/>The last 20 nts of eGFP, reverse complement<\/p><p>5&#8242;-<strong>GGG<\/strong>\u00a0<span style=\"color: #008000;\">GGATTC<\/span>\u00a0<span style=\"color: #ff6600;\">TTTGTATAGTTCATCCATGC<\/span>-3&#8242;<\/p><\/div>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t<div class=\"elementor-column elementor-col-50 elementor-inner-column elementor-element elementor-element-25ea4d8\" data-id=\"25ea4d8\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-0f3b1f7 elementor-widget elementor-widget-image\" data-id=\"0f3b1f7\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t<figure class=\"wp-caption\">\n\t\t\t\t\t\t\t\t\t\t<img width=\"675\" height=\"970\" src=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag-vector-protocol@2x.webp\" class=\"attachment-large size-large\" alt=\"\" loading=\"lazy\" srcset=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag-vector-protocol@2x.webp 675w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag-vector-protocol@2x-209x300.webp 209w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag-vector-protocol@2x-418x600.webp 418w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag-vector-protocol@2x-111x160.webp 111w\" sizes=\"(max-width: 675px) 100vw, 675px\" \/>\t\t\t\t\t\t\t\t\t\t\t<figcaption class=\"widget-image-caption wp-caption-text\">Fig. 4: Descripci\u00f3n general del procedimiento de clonaci\u00f3n para agregar una etiqueta GST a una prote\u00edna de inter\u00e9s.<\/figcaption>\n\t\t\t\t\t\t\t\t\t\t<\/figure>\n\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-e4886ef elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"e4886ef\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-34e8b9b\" data-id=\"34e8b9b\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-c4d584a elementor-widget elementor-widget-text-editor\" data-id=\"c4d584a\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p><strong>Importante:<\/strong> Tenga en cuenta que esta estrategia presentada es solo un ejemplo de c\u00f3mo se puede agregar una etiqueta GST a una prote\u00edna de inter\u00e9s. Otros m\u00e9todos como la recombinaci\u00f3n hom\u00f3loga en levaduras o bacterias tambi\u00e9n son formas posibles de lograr esto.<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-7286b13 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"7286b13\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-f74c86f\" data-id=\"f74c86f\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-c4b647b elementor-widget elementor-widget-spacer\" data-id=\"c4b647b\" data-element_type=\"widget\" data-widget_type=\"spacer.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-spacer\">\n\t\t\t<div class=\"elementor-spacer-inner\"><\/div>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-3bd3828 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"3bd3828\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-1f58a17\" data-id=\"1f58a17\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-97808b7 elementor-widget elementor-widget-text-editor\" data-id=\"97808b7\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Tabla 2: Diferentes vectores pGEX y los sitios de restricci\u00f3n dentro de sus MCS, ordenados en direcci\u00f3n 5&#8242; a 3&#8242;.<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-ca07d26 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"ca07d26\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-83d2482\" data-id=\"83d2482\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-815b434 elementor-widget elementor-widget-text-editor\" data-id=\"815b434\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<table class=\"col-12 responsiv\"><tbody><tr><th>PGEX VECTOR<\/th><th>RESTRICTION SITES IN 5&#8242; TO 3&#8242; DIRECTION<\/th><\/tr><tr><td><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13849.1\" target=\"_blank\" rel=\"noopener\">pGEX-1\u03bbT<\/a><\/td><td>BamHI , EcoRI<\/td><\/tr><tr><td><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13850.1\" target=\"_blank\" rel=\"noopener\">pGEX-2T<\/a><\/td><td>BamHI, SmaI, EcoRI<\/td><\/tr><tr><td><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13851.1\" target=\"_blank\" rel=\"noopener\">pGEX-2TK<\/a><\/td><td>BamHI, SmaI, EcoRI<\/td><\/tr><tr><td><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13853.1\" target=\"_blank\" rel=\"noopener\">pGEX-4T-1<\/a><\/td><td>BamHI, EcoRI, SmaI, SalI, XhoI, NotI<\/td><\/tr><tr><td><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13854.1\" target=\"_blank\" rel=\"noopener\">pGEX-4T-2<\/a><\/td><td>BamHI, EcoRI, SmaI, SalI, XhoI, NotI<\/td><\/tr><tr><td><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13855.1\" target=\"_blank\" rel=\"noopener\">pGEX-4T-3<\/a><\/td><td>BamHI, EcoRI, SmaI, SalI, XhoI, NotI<\/td><\/tr><tr><td><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13852.1\" target=\"_blank\" rel=\"noopener\">pGEX-3X<\/a><\/td><td>BamHI, SmaI, EcoRI,<\/td><\/tr><tr><td><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13856.1\" target=\"_blank\" rel=\"noopener\">pGEX-5X-1<\/a><\/td><td>BamHI, EcoRI, SmaI, SalI, XhoI, NotI<\/td><\/tr><tr><td><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13857.1\" target=\"_blank\" rel=\"noopener\">pGEX-5X-2<\/a><\/td><td>BamHI, EcoRI, SmaI, SalI, XhoI, NotI<\/td><\/tr><tr><td><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13858.1\" target=\"_blank\" rel=\"noopener\">pGEX-5X-3<\/a><\/td><td>BamHI, EcoRI, SmaI, SalI, XhoI, NotI<\/td><\/tr><tr><td><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U78872.1\" target=\"_blank\" rel=\"noopener\">pGEX-6P-1<\/a><\/td><td>BamHI, EcoRI, SmaI SalI, XhoI, NotI<\/td><\/tr><tr><td><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U78873.1\" target=\"_blank\" rel=\"noopener\">pGEX-6P-2<\/a><\/td><td>BamHI, EcoRI, SmaI SalI, XhoI, NotI<\/td><\/tr><tr><td><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U78874.1\" target=\"_blank\" rel=\"noopener\">pGEX-6P-3<\/a><\/td><td>BamHI, EcoRI, SmaI SalI, XhoI, NotI<\/td><\/tr><\/tbody><\/table>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-c66acac elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"c66acac\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-e083460\" data-id=\"e083460\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-ef626a2 elementor-widget-divider--view-line elementor-widget elementor-widget-divider\" data-id=\"ef626a2\" data-element_type=\"widget\" data-widget_type=\"divider.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-divider\">\n\t\t\t<span class=\"elementor-divider-separator\">\n\t\t\t\t\t\t<\/span>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-f79c749 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"f79c749\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-7b775d5\" data-id=\"7b775d5\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<section class=\"elementor-section elementor-inner-section elementor-element elementor-element-bb6c685 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"bb6c685\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-50 elementor-inner-column elementor-element elementor-element-03d600a\" data-id=\"03d600a\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-4c02fcd elementor-widget elementor-widget-text-editor\" data-id=\"4c02fcd\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>\u00bfC\u00f3mo funciona la purificaci\u00f3n de prote\u00ednas con etiqueta GST?<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-91d10ff elementor-widget elementor-widget-text-editor\" data-id=\"91d10ff\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>La purificaci\u00f3n de prote\u00ednas con etiqueta GST es un m\u00e9todo de cromatograf\u00eda de afinidad. Aprovecha la alta afinidad de GST hacia el glutati\u00f3n reducido, su sustrato natural. Para lograr esto, el glutati\u00f3n se une a resina de agarosa o perlas magn\u00e9ticas, seg\u00fan el m\u00e9todo de purificaci\u00f3n preferido. La figura 5 muestra una representaci\u00f3n esquem\u00e1tica de tal cuenta.<\/p><p>El protocolo de purificaci\u00f3n real difiere de un proveedor a otro de productos de afinidad con etiquetas GST. Estas diferencias tambi\u00e9n pueden incluir diferentes composiciones de tampones que solo est\u00e1n optimizadas para sus propias perlas de afinidad GST espec\u00edficas. Por lo tanto, recomendamos encarecidamente no aplicar protocolos para la purificaci\u00f3n de prote\u00ednas con etiqueta GST de un proveedor al producto de otro proveedor.<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-8ce6ddd elementor-widget elementor-widget-text-editor\" data-id=\"8ce6ddd\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Usted puede encontrar los productos\u00a0<a href=\"https:\/\/cube-biotech.com\/products\/protein-purification-products\/gst-tag-affinity-resins-magbeads\/\">GST-tag affinity <\/a>de Cube Biotech en este <a href=\"https:\/\/cube-biotech.com\/products\/protein-purification-products\/gst-tag-affinity-resins-magbeads\/\" target=\"_blank\" rel=\"noopener\">enlace.<\/a>\u00a0<\/p><p>Los protocolos correspondientes se enlistan aqui:<br \/><a href=\"https:\/\/cube-biotech.com\/purification-of-gst-tagged-proteins-using-purecube-glutathione-agarose\">Agarose Resin GST-tag purification protocol:<\/a><br \/><a href=\"https:\/\/cube-biotech.com\/purification-of-gst-tagged-proteins-using-purecube-glutathione-magbeads\">Magnetic Bead GST-tag purification protocol:<\/a><\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t<div class=\"elementor-column elementor-col-50 elementor-inner-column elementor-element elementor-element-457bc78\" data-id=\"457bc78\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-73e6e79 elementor-widget elementor-widget-image\" data-id=\"73e6e79\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<img width=\"568\" height=\"884\" src=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag-interaction-magbeads.jpg\" class=\"attachment-large size-large\" alt=\"\" loading=\"lazy\" srcset=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag-interaction-magbeads.jpg 568w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag-interaction-magbeads-193x300.jpg 193w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag-interaction-magbeads-386x600.jpg 386w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag-interaction-magbeads-103x160.jpg 103w\" sizes=\"(max-width: 568px) 100vw, 568px\" \/>\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-6bc8dcc elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"6bc8dcc\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-55d5378\" data-id=\"55d5378\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-929d892 elementor-widget elementor-widget-spacer\" data-id=\"929d892\" data-element_type=\"widget\" data-widget_type=\"spacer.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-spacer\">\n\t\t\t<div class=\"elementor-spacer-inner\"><\/div>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-085aff5 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"085aff5\" data-element_type=\"section\" data-settings=\"{&quot;background_background&quot;:&quot;gradient&quot;}\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-extended\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-210f5ae\" data-id=\"210f5ae\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<section class=\"elementor-section elementor-inner-section elementor-element elementor-element-8e6599c elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"8e6599c\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-50 elementor-inner-column elementor-element elementor-element-7fe7bf7\" data-id=\"7fe7bf7\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-b37e335 elementor-widget elementor-widget-image\" data-id=\"b37e335\" data-element_type=\"widget\" data-widget_type=\"image.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t\t\t\t\t\t<figure class=\"wp-caption\">\n\t\t\t\t\t\t\t\t\t\t<img width=\"706\" height=\"716\" src=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag-purification-magbeads.jpg\" class=\"attachment-large size-large\" alt=\"\" loading=\"lazy\" srcset=\"https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag-purification-magbeads.jpg 706w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag-purification-magbeads-296x300.jpg 296w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag-purification-magbeads-592x600.jpg 592w, https:\/\/nuevo22.cidsamexico.com\/wp-content\/uploads\/2023\/04\/GST-tag-purification-magbeads-158x160.jpg 158w\" sizes=\"(max-width: 706px) 100vw, 706px\" \/>\t\t\t\t\t\t\t\t\t\t\t<figcaption class=\"widget-image-caption wp-caption-text\">Fig. 6: Overview of the workflow of an exemplary GST-tag protein purification using magnetic beads.<\/figcaption>\n\t\t\t\t\t\t\t\t\t\t<\/figure>\n\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t<div class=\"elementor-column elementor-col-50 elementor-inner-column elementor-element elementor-element-1cb5bbf\" data-id=\"1cb5bbf\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-c3f621c elementor-widget elementor-widget-text-editor\" data-id=\"c3f621c\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Prote\u00edna\u00a0 GST-Tag<br \/>Protocolo de purificaci\u00f3n<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<div class=\"elementor-element elementor-element-0446c7d elementor-widget elementor-widget-text-editor\" data-id=\"0446c7d\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>La purificaci\u00f3n de prote\u00ednas con perlas de afinidad GST se presenta en la figura 5. Estos pasos principales se basan en el procedimiento de purificaci\u00f3n que utiliza perlas magn\u00e9ticas de afinidad con etiqueta GST.<\/p><p><strong>Paso 1:<\/strong> El lisado celular despu\u00e9s de la lisis celular. Todo el proteoma de la c\u00e9lula flota dentro del tamp\u00f3n de lisis. Las prote\u00ednas etiquetadas con GST no est\u00e1n separadas de las otras prote\u00ednas.<\/p><p><strong>Paso 2:<\/strong> Adici\u00f3n de Affinity MagBeads con etiqueta GST al lisado celular. Deben agregarse m\u00e1s MagBeads de las que se sospecha de prote\u00edna etiquetada con GST dentro del lisado celular.<\/p><p><strong>Paso 3:<\/strong> rotaci\u00f3n del cubo\/matraz durante un par de horas para dar a las perlas de glutati\u00f3n tiempo suficiente para unir todas las prote\u00ednas etiquetadas con GST dentro del lisado celular.<\/p><p><strong>Paso 4:<\/strong> las prote\u00ednas marcadas con GST ahora se han unido a MagBeads. Debido a la especificidad de la etiqueta GST con el glutati\u00f3n en las perlas, ninguna otra prote\u00edna deber\u00eda haberse unido.<\/p><p><strong>Paso 5:<\/strong> Aplicaci\u00f3n de una fuerza magn\u00e9tica sobre el lisado celular. Las prote\u00ednas etiquetadas con GST ahora se han unido a las perlas de glutati\u00f3n, mientras que ninguna otra prote\u00edna lo ha hecho.<\/p><p><strong>Paso 6:<\/strong> Eliminaci\u00f3n de la prote\u00edna\/lisado celular no unido. Las prote\u00ednas marcadas con GST ahora est\u00e1n purificadas.<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-c14d2d3 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"c14d2d3\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-a69a961\" data-id=\"a69a961\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-9ea5a6f elementor-widget elementor-widget-text-editor\" data-id=\"9ea5a6f\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Para verificar el \u00e9xito de una purificaci\u00f3n con etiqueta GST, recomendamos realizar un Western Blot que utilice anticuerpos anti-GST para detectar la prote\u00edna purificada. Puede encontrar un protocolo de ejemplo de Western Blot <a href=\"https:\/\/cube-biotech.com\/western-blot-detection-with-gst-tagged-protein\" target=\"_blank\" rel=\"noopener\">aqu\u00ed.<\/a><\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-df8ae90 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"df8ae90\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-e779f1f\" data-id=\"e779f1f\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-520dd4f elementor-widget-divider--view-line elementor-widget elementor-widget-divider\" data-id=\"520dd4f\" data-element_type=\"widget\" data-widget_type=\"divider.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-divider\">\n\t\t\t<span class=\"elementor-divider-separator\">\n\t\t\t\t\t\t<\/span>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-55b1652 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"55b1652\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-745ab56\" data-id=\"745ab56\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-fc77483 elementor-widget elementor-widget-text-editor\" data-id=\"fc77483\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>\u00bfC\u00f3mo eliminar una etiqueta GST?<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-ac54a39 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"ac54a39\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-94f26dd\" data-id=\"94f26dd\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-1721007 elementor-widget elementor-widget-text-editor\" data-id=\"1721007\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Con 26 kilodaltons (kDa), la etiqueta GST es una de las etiquetas de mayor afinidad que existen. Como de hecho es una prote\u00edna completa, puede incluso superar algunas prote\u00ednas a las que se une. Por supuesto, esto crea la preocupaci\u00f3n v\u00e1lida de que una uni\u00f3n artificial de este tama\u00f1o a una prote\u00edna pueda interferir con la funcionalidad de dicha prote\u00edna. Por lo tanto, surge la pregunta de si es posible eliminar una etiqueta GST despu\u00e9s de la purificaci\u00f3n de la prote\u00edna para garantizar que no obstaculice la prote\u00edna purificada de ninguna manera.<\/p><p>Esta preocupaci\u00f3n ya estaba en la mente de los creadores de la serie de vectores pGEX, que ya mencionamos en la tabla 2. Al crear una prote\u00edna de fusi\u00f3n de etiqueta GST de la manera descrita anteriormente en el cap\u00edtulo &#8220;C\u00f3mo agregar\/clonar una etiqueta GST en un pl\u00e1smido&#8221;, tiene la opci\u00f3n de eliminar esta etiqueta GST m\u00e1s adelante.<\/p><p>La respuesta es agregar un sitio de escisi\u00f3n de prote\u00edna espec\u00edfico en la direcci\u00f3n 5&#8242; del MCS del pl\u00e1smido. De esta manera, la etiqueta GST N-terminal se puede escindir, incluso despu\u00e9s de la purificaci\u00f3n. Esto tambi\u00e9n se puede usar como un m\u00e9todo de eluci\u00f3n durante la purificaci\u00f3n en lugar del tamp\u00f3n de eluci\u00f3n que se menciona en nuestros protocolos de purificaci\u00f3n con resina de agarosa y perlas magn\u00e9ticas.<\/p><p>Dado que este m\u00e9todo incluye una peque\u00f1a posibilidad de que su prote\u00edna de inter\u00e9s incluya un sitio de escisi\u00f3n de proteasa por coincidencia, hay tres opciones diferentes de sitios de escisi\u00f3n de prote\u00ednas que ya est\u00e1n presentes en los vectores pGEX.<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-a2ede65 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"a2ede65\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-f843dd8\" data-id=\"f843dd8\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-7986d05 elementor-widget elementor-widget-spacer\" data-id=\"7986d05\" data-element_type=\"widget\" data-widget_type=\"spacer.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-spacer\">\n\t\t\t<div class=\"elementor-spacer-inner\"><\/div>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-c5cfa9d elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"c5cfa9d\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-20d71f6\" data-id=\"20d71f6\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-12d568f elementor-widget elementor-widget-text-editor\" data-id=\"12d568f\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Tabla 3: Diferentes vectores pGEX y su sitio de escisi\u00f3n de proteasa incluido.<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-72c6a26 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"72c6a26\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-d65031e\" data-id=\"d65031e\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-cc4c145 elementor-widget elementor-widget-text-editor\" data-id=\"cc4c145\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<div><table class=\"col-12 cube_table\"><tbody><tr><th>PGEX VECTOR<\/th><th>PROTEASE CLEAVAGE SITE IN 5&#8242; OF THE MCS<\/th><\/tr><tr><td><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13849.1\" target=\"_blank\" rel=\"noopener\">pGEX-1\u03bbT<\/a>;\u00a0<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13850.1\" target=\"_blank\" rel=\"noopener\">pGEX-2T<\/a>;<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13851.1\" target=\"_blank\" rel=\"noopener\">\u00a0pGEX-2TK<\/a>;<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13853.1\" target=\"_blank\" rel=\"noopener\">\u00a0pGEX-4T-1<\/a>;<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13854.1\" target=\"_blank\" rel=\"noopener\">\u00a0pGEX-4T-2<\/a>;\u00a0<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13855.1\" target=\"_blank\" rel=\"noopener\">pGEX-4T-3<\/a><\/td><td>Thrombin<\/td><\/tr><tr><td><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13852.1\" target=\"_blank\" rel=\"noopener\">pGEX-3X<\/a>;<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13856.1\" target=\"_blank\" rel=\"noopener\">\u00a0pGEX-5X-1<\/a>;<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13857.1\" target=\"_blank\" rel=\"noopener\">\u00a0pGEX-5X-2<\/a>;<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U13858.1\" target=\"_blank\" rel=\"noopener\">\u00a0pGEX-5X-3<\/a><\/td><td>Factor Xa<\/td><\/tr><tr><td><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U78872.1\" target=\"_blank\" rel=\"noopener\">pGEX-6P-1<\/a>;<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U78873.1\" target=\"_blank\" rel=\"noopener\">\u00a0pGEX-6P-2<\/a>;\u00a0<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/nuccore\/U78874.1\" target=\"_blank\" rel=\"noopener\">pGEX-6P-3<\/a><\/td><td>PreScission Protease<\/td><\/tr><\/tbody><\/table><\/div>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-04f0ab0 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"04f0ab0\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-0a539a8\" data-id=\"0a539a8\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-2a0e548 elementor-widget elementor-widget-spacer\" data-id=\"2a0e548\" data-element_type=\"widget\" data-widget_type=\"spacer.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-spacer\">\n\t\t\t<div class=\"elementor-spacer-inner\"><\/div>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-3f7c556 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"3f7c556\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-2b0e1c4\" data-id=\"2b0e1c4\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-9f563a0 elementor-widget elementor-widget-text-editor\" data-id=\"9f563a0\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Ejemplo de Protocolo &#8211; Eliminaci\u00f3n de GST-Tag\u00a0<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-ac4e280 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"ac4e280\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-dffe8ef\" data-id=\"dffe8ef\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-1bee519 elementor-widget elementor-widget-text-editor\" data-id=\"1bee519\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p><strong>Nota:<\/strong> Este protocolo describe c\u00f3mo escindir una etiqueta GST utilizando el sitio de escisi\u00f3n de trombina de prote\u00ednas de fusi\u00f3n GST eluidas libres.<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-4412491 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"4412491\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-68822ff\" data-id=\"68822ff\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-8dfea47 elementor-widget elementor-widget-spacer\" data-id=\"8dfea47\" data-element_type=\"widget\" data-widget_type=\"spacer.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-spacer\">\n\t\t\t<div class=\"elementor-spacer-inner\"><\/div>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-99521f3 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"99521f3\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-9b1bf63\" data-id=\"9b1bf63\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-657d571 elementor-widget elementor-widget-text-editor\" data-id=\"657d571\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Tabla 4: Reactivos necesarios para la escisi\u00f3n de trombina<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-8922cd8 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"8922cd8\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-5512cce\" data-id=\"5512cce\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-559af90 elementor-widget elementor-widget-text-editor\" data-id=\"559af90\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<div class=\"col-12\"><table class=\"responsiv\"><tbody><tr><th>REAGENT<\/th><th>AMOUNT<\/th><\/tr><tr><td>Thrombin<\/td><td><em>Depends on the supplier:<\/em>\u00a03-10 U of thrombin can cleave 1 mg of fusion protein at RT overnight on average. Still, read the supplier&#8217;s notes.<\/td><\/tr><tr><td>1 X PBS buffer<\/td><td>140mM NaCl; 2.7mM KCl; 10mM Na<sub>2<\/sub>HPO<sub>4<\/sub>; 1.8mM KH<sub>2<\/sub>PO<sub>4<\/sub>; pH 7.3<\/td><\/tr><tr><td>PMSF<\/td><td>Enough for a 1 mM concentration in your protein solution<\/td><\/tr><tr><td>AEBSF<\/td><td><em>Alternative to PMFS;<\/em>\u00a0Enough for a 1mM concentration in your protein solution<\/td><\/tr><\/tbody><\/table><\/div>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-88ea920 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"88ea920\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-84fd85f\" data-id=\"84fd85f\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-48f106f elementor-widget elementor-widget-text-editor\" data-id=\"48f106f\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Procedimiento (version general)<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-669623f elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"669623f\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-60d97e9\" data-id=\"60d97e9\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-132fc56 elementor-widget elementor-widget-text-editor\" data-id=\"132fc56\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>&#8211; A\u00f1adir al eluato o columna de purificaci\u00f3n 10 unidades de escisi\u00f3n de trombina ~10 \u00b5l por miligramo de prote\u00edna de fusi\u00f3n GST<br \/>&#8211; Mezcle suavemente e incube la mezcla a temperatura ambiente (22\u00b0C) durante 2 a 16 horas.<br \/>&#8211; Despu\u00e9s de la digesti\u00f3n, puede detener la reacci\u00f3n agregando 1 mM de PMSF o ABESF<br \/>&#8211; Verifique los resultados a trav\u00e9s de la p\u00e1gina SDS\u00a0 \u00f3<br \/>&#8211; Separe directamente los productos de escisi\u00f3n mediante cromatograf\u00eda SDS (por ejemplo, cromatograf\u00eda de exclusi\u00f3n por tama\u00f1o)<\/p><p><br \/>Si ha encontrado sus condiciones de divisi\u00f3n satisfactorias, puede escalar el procedimiento de forma lineal.<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-28a0155 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"28a0155\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-cf9682e\" data-id=\"cf9682e\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-e2c653a elementor-widget elementor-widget-text-editor\" data-id=\"e2c653a\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Tip: Puede incubar una cantidad suficiente de su prote\u00edna etiquetada con GST para ver en geles SDS-Page con 0,01 a 0,06 unidades de trombina y diferentes tiempos de digesti\u00f3n (por ejemplo, 2 h, 4 h, 6 h, etc.). Detenga la reacci\u00f3n como se describe y almacene las al\u00edcuotas a -20\u00b0C hasta su uso.<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-0640952 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"0640952\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-65e3942\" data-id=\"65e3942\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-bb7bfd4 elementor-widget-divider--view-line elementor-widget elementor-widget-divider\" data-id=\"bb7bfd4\" data-element_type=\"widget\" data-widget_type=\"divider.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-divider\">\n\t\t\t<span class=\"elementor-divider-separator\">\n\t\t\t\t\t\t<\/span>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-a62a01b elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"a62a01b\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-1c9f554\" data-id=\"1c9f554\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-f6c0165 elementor-widget elementor-widget-text-editor\" data-id=\"f6c0165\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<p>Referencias<\/p>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-9f00e44 elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"9f00e44\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-991f4e9\" data-id=\"991f4e9\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-ce49593 elementor-widget elementor-widget-text-editor\" data-id=\"ce49593\" data-element_type=\"widget\" data-widget_type=\"text-editor.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t\t\t<ol><li>Hayes, J. D., Flanagan, J. U. &amp; Jowsey, I. R. GLUTATHIONE TRANSFERASES.\u00a0<em>Annu. Rev. Pharmacol. Toxicol.<\/em>\u00a0<strong>45<\/strong>, 51\u201388 (2004).<\/li><li>Allocati, N., Federici, L., Masulli, M. &amp; Di Ilio, C. Glutathione transferases in bacteria.\u00a0<em>The FEBS Journal<\/em>\u00a0<strong>276<\/strong>, 58\u201375 (2009).<\/li><li>Oakley, A. Glutathione transferases: A structural perspective.\u00a0<em>Drug metabolism reviews<\/em>\u00a0<strong>43<\/strong>, 138\u201351 (2011).<\/li><li>Smith, D. B.\u00a0<em>et al.<\/em>\u00a0Mr 26,000 antigen of Schistosoma japonicum recognized by resistant WEHI 129\/J mice is a parasite glutathione S-transferase.\u00a0<em>Proc Natl Acad Sci U S A<\/em>\u00a0<strong>83<\/strong>, 8703\u20138707 (1986).<\/li><li>Terpe, K. Overview of tag protein fusions: from molecular and biochemical fundamentals to commercial systems.\u00a0<em>Applied Microbiology and Biotechnology<\/em>\u00a0<strong>60<\/strong>, 523\u2013533 (2003).<\/li><li>Rudert, F.\u00a0<em>et al.<\/em>\u00a0pLEF, a novel vector for expression of glutathione S-transferase fusion proteins in mammalian cells.\u00a0<em>Gene<\/em>\u00a0<strong>169<\/strong>, 281\u2013282 (1996).<\/li><li>Harper, S. &amp; Speicher, D. W. Purification of proteins fused to glutathione S-transferase.\u00a0<em>Methods Mol Biol<\/em>\u00a0<strong>681<\/strong>, 259\u2013280 (2011).<\/li><li>Smith, D. B. &amp; Johnson, K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase.\u00a0<em>Gene<\/em>\u00a0<strong>67<\/strong>, 31\u201340 (1988).<\/li><li>Lu, Q., Bauer, J. C. &amp; Greener, A. Using Schizosaccharomyces pombe as a host for expression and purification of eukaryotic proteins.\u00a0<em>Gene<\/em>\u00a0<strong>200<\/strong>, 135\u2013144 (1997).<\/li><\/ol>\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t<section class=\"elementor-section elementor-top-section elementor-element elementor-element-471bedf elementor-section-boxed elementor-section-height-default elementor-section-height-default\" data-id=\"471bedf\" data-element_type=\"section\">\n\t\t\t\t\t\t<div class=\"elementor-container elementor-column-gap-default\">\n\t\t\t\t\t<div class=\"elementor-column elementor-col-100 elementor-top-column elementor-element elementor-element-777e388\" data-id=\"777e388\" data-element_type=\"column\">\n\t\t\t<div class=\"elementor-widget-wrap elementor-element-populated\">\n\t\t\t\t\t\t\t\t<div class=\"elementor-element elementor-element-f25ac16 elementor-widget elementor-widget-spacer\" data-id=\"f25ac16\" data-element_type=\"widget\" data-widget_type=\"spacer.default\">\n\t\t\t\t<div class=\"elementor-widget-container\">\n\t\t\t\t\t<div class=\"elementor-spacer\">\n\t\t\t<div class=\"elementor-spacer-inner\"><\/div>\n\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\t\t<\/div>\n\t\t<\/div>\n\t\t\t\t\t\t\t<\/div>\n\t\t<\/section>\n\t\t\t\t\t\t\t<\/div>\n\t\t","protected":false},"excerpt":{"rendered":"<p>Como una proteina ayuda a purificar a otra proteina\u00a0 Figura 1: La etiqueta GST y sus caracteristicas \u00bfQu\u00e9 es la etiqueta GST? Existen m\u00faltiples etiquetas de afinidad de prote\u00ednas que se utilizan para la purificaci\u00f3n de prote\u00ednas y la etiqueta GST es una de ellas. GST es la abreviatura de glutati\u00f3n-S-transferasa y se refiere a [&hellip;]<\/p>\n","protected":false},"author":2,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"om_disable_all_campaigns":false,"_monsterinsights_skip_tracking":false,"_monsterinsights_sitenote_active":false,"_monsterinsights_sitenote_note":"","_monsterinsights_sitenote_category":0},"aioseo_notices":[],"_links":{"self":[{"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/pages\/2445"}],"collection":[{"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/comments?post=2445"}],"version-history":[{"count":43,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/pages\/2445\/revisions"}],"predecessor-version":[{"id":11189,"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/pages\/2445\/revisions\/11189"}],"wp:attachment":[{"href":"https:\/\/nuevo22.cidsamexico.com\/index.php\/wp-json\/wp\/v2\/media?parent=2445"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}