<?xml version="1.0" encoding="ISO-8859-1"?><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance">
<front>
<journal-meta>
<journal-id>0123-3475</journal-id>
<journal-title><![CDATA[Revista Colombiana de Biotecnología]]></journal-title>
<abbrev-journal-title><![CDATA[Rev. colomb. biotecnol]]></abbrev-journal-title>
<issn>0123-3475</issn>
<publisher>
<publisher-name><![CDATA[Instituto de Biotecnología, Universidad Nacional de Colombia]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S0123-34752023000100057</article-id>
<article-id pub-id-type="doi">10.15446/rev.colomb.biote.v25n1.91675</article-id>
<title-group>
<article-title xml:lang="es"><![CDATA[Obtención de la proteína verde fluorescente recombinante y su anticuerpo policlonal Igy]]></article-title>
<article-title xml:lang="en"><![CDATA[Obtaining the recombinant green fluorescent protein and its IgY polyclonal antibody]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Gómez Osorio]]></surname>
<given-names><![CDATA[Valentina]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[González Rodríguez]]></surname>
<given-names><![CDATA[Sebastián]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Contreras Rodríguez]]></surname>
<given-names><![CDATA[Luis Ernesto]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Díaz]]></surname>
<given-names><![CDATA[Gonzalo Jair]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Ramírez Hernández]]></surname>
<given-names><![CDATA[María Helena]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Universidad Nacional de Colombia Facultad de Ciencias ]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
<country>Colombia</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>06</month>
<year>2023</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>06</month>
<year>2023</year>
</pub-date>
<volume>25</volume>
<numero>1</numero>
<fpage>57</fpage>
<lpage>68</lpage>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_arttext&amp;pid=S0123-34752023000100057&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_abstract&amp;pid=S0123-34752023000100057&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.org.co/scielo.php?script=sci_pdf&amp;pid=S0123-34752023000100057&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="es"><p><![CDATA[RESUMEN La Proteína Verde Fluorescente (Green Fluorescent Protein, GFP) es ampliamente utilizada en ensayos in vivo e in vitro. Se han generado múltiples variantes de esta proteína para diversificar sus características, como la GFP-enhancer (EGFP) que emite una señal de fluorescencia 35 veces mayor en comparación con la proteína silvestre, siendo implementada como proteína fusión en estudios de localización y estabilidad estructural, entre otros. La detección de esta proteína y sus variantes puede ser directa o indirecta, mediante el uso de anticuerpos anti-GFP. Aunque el uso de GFP es generalizado y de evidente utilidad en investigación y en docencia, los insumos para su estudio exhiben un alto costo dado que deben ser importados, constituyendo un recurso limitado en Colombia. El presente trabajo reporta la clonación y expresión de la proteína recombinante 6xHisEGFP, cuya purificación se completó a partir de la fracción soluble e insoluble del sistema heterólogo Escherichia coli mediante cromatografía de afinidad a metales inmovilizados y electroforesis preparativa, respectivamente. La proteína purificada se implementó como antígeno para la producción de anticuerpos policlonales aviares (IgY) contra la EGFP, los cuales se obtuvieron desde los huevos colectados y el suero de las sangrías de las gallinas inmunizadas. En este sentido, la estrategia metodológica planteada constituye un avance en el desarrollo de un sistema biotecnológico para la producción nacional de herramientas moleculares como los anticuerpos policlonales aviares a bajo costo.]]></p></abstract>
<abstract abstract-type="short" xml:lang="en"><p><![CDATA[ABSTRACT Green Fluorescent Protein (GFP) is widely used in in vivo and in vitro assays. Multiple variants of this protein have been generated to diversify its characteristics, such as the enhancer GFP (EGFP) that emits a 35-fold higher fluorescence signal compared to the wild-type protein, being implemented as a fusion reporter in localization and structural stability studies, among others. Detection of this protein can be direct or indirect, fusing anti-GFP antibodies. Although the use of GFP is generalized and of evident utility in research and teaching, the molecular tools for its study exhibit a high cost since they must be imported, constituting a limited resource in Colombia. This work reports the cloning and expression of the recombinant protein 6xHisEGFP, which purification was completed from the soluble and insoluble fraction of the heterologous Escherichia coli system by immobilized metal affinity chromatography and preparative SDS-PAGE, respectively. The purified protein was implemented as an antigen to produce avian polyclonal antibodies (IgY) against EGFP, which were obtained from collected eggs and blood serum from immunized hens. In this sense, the proposed methodological strategy constitutes an advance in the development of a biotechnological system for the national production of molecular tools such as avian polyclonal antibodies at low-cost.]]></p></abstract>
<kwd-group>
<kwd lng="es"><![CDATA[Biotecnología]]></kwd>
<kwd lng="es"><![CDATA[EGFP]]></kwd>
<kwd lng="es"><![CDATA[Proteínas recombinantes]]></kwd>
<kwd lng="en"><![CDATA[Biotechnology]]></kwd>
<kwd lng="en"><![CDATA[EGFP]]></kwd>
<kwd lng="en"><![CDATA[Recombinant proteins]]></kwd>
</kwd-group>
</article-meta>
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