<?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>S0379-39822020000200027</article-id>
<article-id pub-id-type="doi">10.18845/tm.v33i1.5017</article-id>
<title-group>
<article-title xml:lang="en"><![CDATA[Alpha-tubulin promoter from Chlorella vulgaris allows genetic transformation of green coccoid microalga]]></article-title>
<article-title xml:lang="es"><![CDATA[El promotor de la alfa-tubulina de Chlorella vulgaris permite la transformación genética de la microalga verde tipo cocoide]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Fernández-Rodríguez]]></surname>
<given-names><![CDATA[Raquel]]></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 contrib-type="author">
<name>
<surname><![CDATA[Guerrero-Barrantes]]></surname>
<given-names><![CDATA[Maritza]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Gómez-Espinoza]]></surname>
<given-names><![CDATA[Olman]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Instituto Tecnológico de Costa Rica Escuela de Biología 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 Escuela de Biología 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 Escuela de Biología 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 Escuela de Biología 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>06</month>
<year>2020</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>06</month>
<year>2020</year>
</pub-date>
<volume>33</volume>
<numero>2</numero>
<fpage>27</fpage>
<lpage>36</lpage>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://www.scielo.sa.cr/scielo.php?script=sci_arttext&amp;pid=S0379-39822020000200027&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.sa.cr/scielo.php?script=sci_abstract&amp;pid=S0379-39822020000200027&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.sa.cr/scielo.php?script=sci_pdf&amp;pid=S0379-39822020000200027&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="en"><p><![CDATA[Abstract Microalgae have become a feasible platform for high-value recombinant protein production. Although diverse sets of genetic tools for microalgae transformation have been developed, some critical elements, such as endogenous promoters, need to be improved for increasing transgenic expression levels after genetic transformation. This work aimed to evaluate a sequence from the 5&#8242; upstream region of the alpha-tubulin gene from Chlorella vulgaris as a promoter for the expression of an antibiotic resistant gene in Chlorella sorokiniana. Using in silico analysis it was possible to identify a proximal promoter corresponding to 281 nucleotides upstream of the ATG start codon, which possessed 9 potential cis-regulatory elements, including TATA Box and CAAT Box. The proximal promoter sequence was used to drive the expression of a streptomycinresistance gene (aada), previous optimization of its codon usage. The codon optimization of the aada sequence allowed investigators to obtain a GC content of 69,8% (compared to 53% of the original sequence), which increases its similarity with Chlorella sp. genomes (67,2%). Both genetic elements were cloned into the pUC57-Kan vector and transformed into C. sorokiniana cells by electroporation. The microalgae transgenic colonies were identified through culture in selective medium and PCR. Our results proved the capacity of the Chlorella vulgaris alpha-tubulin promoter to express a foreign antibiotic-resistance gene in C. sorokiniana cells. Research of endogenous promoting sequences is essential in order to accomplish an efficient heterologous gene expression, especially in microalgae used for industrial production like those from Chlorella genus.]]></p></abstract>
<abstract abstract-type="short" xml:lang="es"><p><![CDATA[Resumen Las microalgas se han convertido en una plataforma viable para la producción de proteínas recombinantes de alto valor. Aunque se han desarrollado diversos conjuntos de herramientas genéticas para la transformación de microalgas, algunos elementos críticos, como los promotores endógenos, deben mejorarse para aumentar los niveles de expresión génica posterior al proceso de transformación. Para contribuir a ello, el presente estudio tuvo como objetivo evaluar una secuencia de la región 5&#8217; del gen alfa-tubulina de Chlorella vulgaris, como promotor para la expresión de un gen resistencia a antibiótico en Chlorella sorokiniana. Mediante análisis in silico se logró identificar un promotor proximal correspondiente a 281 nucleótidos corriente arriba del codón de inicio ATG, el cual poseía 9 potenciales elementos reguladores en cis, incluyendo Caja TATA y Caja CAAT. La secuencia promotora proximal se usó para dirigir la expresión de un gen de resistencia a la estreptomicina (aada), previa optimización de su uso de codones. La optimización de codones de la secuencia aada permitió obtener un contenido de GC del 69,8% (en comparación con el 53% del gen original), lo que aumenta su similitud con los genomas de Chlorella sp. (67,2%). Ambos elementos genéticos se clonaron en el vector pUC57-Kan y se transformaron en células de C. sorokiniana mediante electroporación. Las colonias de microalgas transgénicas fueron identificadas a través de cultivo en medio selectivo y PCR. Nuestros resultados demostraron la capacidad del promotor alfa-tubulina de Chlorella vulgaris para expresar un gen heterólogo de resistencia a antibiótico en células de C. sorokiniana. La investigación de secuencias promotoras endógenas es esencial para lograr una expresión génica heteróloga eficiente, especialmente en microalgas utilizadas para la producción industrial como las del género Chlorella.]]></p></abstract>
<kwd-group>
<kwd lng="es"><![CDATA[Electroporación]]></kwd>
<kwd lng="es"><![CDATA[kazusa]]></kwd>
<kwd lng="es"><![CDATA[estreptomicina]]></kwd>
<kwd lng="es"><![CDATA[transgénico]]></kwd>
<kwd lng="es"><![CDATA[Caja TATA]]></kwd>
<kwd lng="en"><![CDATA[Electroporation]]></kwd>
<kwd lng="en"><![CDATA[kazusa]]></kwd>
<kwd lng="en"><![CDATA[streptomycin]]></kwd>
<kwd lng="en"><![CDATA[transgenic]]></kwd>
<kwd lng="en"><![CDATA[TATA-box]]></kwd>
</kwd-group>
</article-meta>
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