<?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>0379-3982</journal-id>
<journal-title><![CDATA[Revista Tecnología en Marcha]]></journal-title>
<abbrev-journal-title><![CDATA[Tecnología en Marcha]]></abbrev-journal-title>
<issn>0379-3982</issn>
<publisher>
<publisher-name><![CDATA[Instituto Tecnológico de Costa Rica]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S0379-39822019000100077</article-id>
<article-id pub-id-type="doi">10.8845/tm.v32.i1.4120</article-id>
<title-group>
<article-title xml:lang="en"><![CDATA[Establecimiento de un protocolo de transformación transitoria y estable de Nicotiana tabacum mediada por Agrobacterium tumefaciens]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Méndez-Muñoz]]></surname>
<given-names><![CDATA[Laura]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Campos-Delgado]]></surname>
<given-names><![CDATA[Sofia]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Obando-Coronado]]></surname>
<given-names><![CDATA[Mariel]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Garro-Monge]]></surname>
<given-names><![CDATA[Giovanni]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Instituto Tecnológico de Costa Rica Centro de Investigación en Biotecnología ]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
<country>Costa Rica</country>
</aff>
<aff id="Af2">
<institution><![CDATA[,Instituto Tecnológico de Costa Rica Centro de Investigación en Biotecnología ]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
<country>Costa Rica</country>
</aff>
<aff id="Af3">
<institution><![CDATA[,Instituto Tecnológico de Costa Rica Centro de Investigación en Biotecnología ]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
<country>Costa Rica</country>
</aff>
<aff id="Af4">
<institution><![CDATA[,Instituto Tecnológico de Costa Rica Centro de Investigación en Biotecnología ]]></institution>
<addr-line><![CDATA[ ]]></addr-line>
<country>Costa Rica</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>03</month>
<year>2019</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>03</month>
<year>2019</year>
</pub-date>
<volume>32</volume>
<numero>1</numero>
<fpage>77</fpage>
<lpage>91</lpage>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://www.scielo.sa.cr/scielo.php?script=sci_arttext&amp;pid=S0379-39822019000100077&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.sa.cr/scielo.php?script=sci_abstract&amp;pid=S0379-39822019000100077&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.sa.cr/scielo.php?script=sci_pdf&amp;pid=S0379-39822019000100077&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="en"><p><![CDATA[Abstract Biotechnological tools such as genetic engineering and molecular biology are very advanced, allowing the development of systems based on the use of plant cells for the production of recombinant proteins, which allows the development of platforms for biopharmaceuticals, enzymes and secondary metabolites production. Transient and stable transformation, allow production of recombinant proteins. In this work a stable transformation of N. tabacum callus from the BY-2 commercial line and a wild one (T1) was performed. In addition, agroinfiltration assays were carried out using two techniques: by syringe and vacuum. In both cases, the transformation was mediated by Agrobacterium tumefaciens LBA4404 which carried the vector pCAMBIA 1303, containing the GFP and GUS marker genes, and a hygromycin resistance gene. The expression of green fluorescent protein was evaluated by fluorescence microscopy. Expression of the hygromycin and &#946;-glucuronidase resistance genes were evaluated by PCR. As a result, high GFP expression and good transformation rates were obtained.]]></p></abstract>
<abstract abstract-type="short" xml:lang="es"><p><![CDATA[Resumen Actualmente las herramientas biotecnológicas como la ingeniería genética y la biología molecular se encuentran muy avanzadas, permitiendo el desarrollo de sistemas que se basan en el uso de células vegetales para la producción de proteínas recombinantes, lo cual permite el desarrollo de plataformas para la elaboración de biofármacos, enzimas y metabolitos secundarios. El uso de metodologías para transformación transitoria y estable, permiten la producción de proteínas recombinantes de interés. En este trabajo se realizó una transformación estable de callo de N. tabacum de la línea comercial BY-2 y una silvestre (T1). Además se realizaron ensayos de agroinfiltración por la técnica al vacío y con la técnica por jeringa. En ambos casos, la transformación fue mediada por Agrobacterium tumefaciens LBA4404 la cual poseía el vector pCAMBIA 1303, conteniendo los genes marcadores GFP y GUS, y un gen de resistencia a Higromicina. Se evaluó la expresión de la proteína verde fluorescente por medio de microscopía de fluorescencia. La expresión de los genes de resistencia a Higromicina y &#946;-glucoronidasa se evaluaron por medio de PCR convencional. Se obtuvieron porcentajes de transformación aceptables, en conjunto con una alta expresión de la proteína verde fluorescente.]]></p></abstract>
<kwd-group>
<kwd lng="es"><![CDATA[Agroinfiltración]]></kwd>
<kwd lng="es"><![CDATA[agroinfección]]></kwd>
<kwd lng="es"><![CDATA[fluorescencia]]></kwd>
<kwd lng="es"><![CDATA[BY-2]]></kwd>
<kwd lng="es"><![CDATA[pCAMBIA 1303]]></kwd>
<kwd lng="es"><![CDATA[acetosiringona]]></kwd>
<kwd lng="es"><![CDATA[Higromicina]]></kwd>
<kwd lng="es"><![CDATA[&#946;-glucoronidasa]]></kwd>
<kwd lng="en"><![CDATA[Agroinfiltration]]></kwd>
<kwd lng="en"><![CDATA[agroinfection]]></kwd>
<kwd lng="en"><![CDATA[fluorescence]]></kwd>
<kwd lng="en"><![CDATA[BY-2]]></kwd>
<kwd lng="en"><![CDATA[pCAMBIA 1303]]></kwd>
<kwd lng="en"><![CDATA[acetosyringone]]></kwd>
<kwd lng="en"><![CDATA[hygromycin]]></kwd>
<kwd lng="en"><![CDATA[&#946;-glucuronidase]]></kwd>
</kwd-group>
</article-meta>
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