<?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>0375-0760</journal-id>
<journal-title><![CDATA[Revista Cubana de Medicina Tropical]]></journal-title>
<abbrev-journal-title><![CDATA[Rev Cubana Med Trop]]></abbrev-journal-title>
<issn>0375-0760</issn>
<publisher>
<publisher-name><![CDATA[Centro Nacional de Información de Ciencias Médicas]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S0375-07602019000200002</article-id>
<title-group>
<article-title xml:lang="en"><![CDATA[Cloning and expression in Escherichia coli of the full-length and deletion variants of a human papillomavirus 18 L1 gene isolated from a Cuban patient]]></article-title>
<article-title xml:lang="es"><![CDATA[Clonación y expresión en Escherichia coli de un gen L1 completo del virus del papiloma humano 18 aislado de una paciente cubana y variantes delecionadas]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Pimienta]]></surname>
<given-names><![CDATA[Elsa]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Rodríguez]]></surname>
<given-names><![CDATA[Sandra]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Fando]]></surname>
<given-names><![CDATA[Rafael]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Serrano]]></surname>
<given-names><![CDATA[Yunier]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Ortega]]></surname>
<given-names><![CDATA[Darién]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Palenzuela]]></surname>
<given-names><![CDATA[Ariel]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Marrero]]></surname>
<given-names><![CDATA[Karen]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,National Center for Scientific Research Development &amp; Innovation Biological Products Research Unit, Direction of Research]]></institution>
<addr-line><![CDATA[La Habana ]]></addr-line>
<country>Cuba</country>
</aff>
<aff id="Af2">
<institution><![CDATA[,Immunoassay Center Laboratory of Infectious Disease ]]></institution>
<addr-line><![CDATA[La Habana ]]></addr-line>
<country>Cuba</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>08</month>
<year>2019</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>08</month>
<year>2019</year>
</pub-date>
<volume>71</volume>
<numero>2</numero>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_arttext&amp;pid=S0375-07602019000200002&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_abstract&amp;pid=S0375-07602019000200002&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://scielo.sld.cu/scielo.php?script=sci_pdf&amp;pid=S0375-07602019000200002&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="en"><p><![CDATA[ABSTRACT  Introduction:  Human papillomavirus (HPV) vaccines are based on the L1 major capsid protein.  Objectives:  To clone the HPV-18 L1 gene from a Cuban female HPV-18-infected patient and to express the full-length and deletion variants of the cloned HPV-18 L1 gene in Escherichia coli.  Methods:  The full-length HPV-18 L1 gene was PCR-amplified from total DNA isolated from a Cuban patient, cloned and finally subcloned into the E. coli expression vector pET26b. Three deletion mutants were constructed, which encode truncated proteins lacking 30 amino acids at the C-terminus in combination with 5, 6 or none deleted residue at the N-terminus. Production of L1 proteins in E. coli BL21(DE3) and E. coli SHuffle T7 was assessed by SDS-PAGE and Western blotting.  Results:  The cloned HPV-18 L1 gene was 99.9 % similar to the African variant EF202152 and probably shares a common origin with the B lineage of genotype 18. The three truncated variants of HPV-18 L1 were produced at higher levels than the full-length HPV-18 L1 protein, attaining higher levels in E. coli BL21(DE3) and higher solubility in E. coli SHuffle. The C-terminus-only truncated variant, L1&#8710;C30, was produced at similar levels to the HPV-18 L1s truncated at both termini. E. coli SHuffle produced about three times more amounts of L1&#8710;C30 when grown under autoinduction conditions with respect to conventional induction and thus, amounts were comparable to those obtained in E. coli BL21(DE3) under conventional induction.  Conclusions:  Truncation of thirty amino acid residues at the carboxy-terminus of the HPV-18 L1 made a major contribution to the production and solubility of this wild-type protein in E. coli. This is the first report about soluble production of HPV-18 L1 protein in an E. coli SHuffle strain. However, higher amounts of L1 are needed to scale-up its production for developing an HPV vaccine candidate.]]></p></abstract>
<abstract abstract-type="short" xml:lang="es"><p><![CDATA[RESUMEN  Introducción:  Las vacunas contra el virus del papiloma humano (VPH) se fundamentan en la proteína principal de la cápsida L1.  Objetivo:  Clonar el gen L1 del VPH-18 a partir de una paciente cubana infectada con VPH-18 y expresar las variantes de longitud completa y delecionadas del gen L1 del VPH-18 en Escherichia coli.  Métodos:  El gen L1 del VPH-18 de longitud completa se amplificó por PCR a partir de ADN total aislado de un paciente cubana, se clonó y finalmente se subclonó en el vector de expresión de E. coli pET26b. Se construyeron tres mutantes de deleción, que codifican para proteínas truncadas que carecen de 30 aminoácidos por el extremo carboxilo, en combinación con 5, 6 o ningún residuo delecionado por el extremo amino. La producción de las proteínas L1 en E. coli BL21(DE3) y E. coli SHuffle T7 se evaluó mediante SDS-PAGE y Western blot.  Resultados:  El gen L1 del VPH-18 clonado fue 99.9 % similar a la variante africana EF202152 y probablemente comparte un origen común con el linaje B del genotipo 18. Las tres variantes truncadas de la proteína L1 del VPH-18 se produjeron a mayores niveles que la proteína L1 del VPH-18 de longitud completa, alcanzando mayores niveles en E. coli BL21(DE3) y mayor solubilidad en E. coli SHuffle. La variante truncada solo por el extremo carboxilo, L1(C30, se produjo a niveles similares a las proteínas L1 del VPH-18 truncadas por ambos extremos. E. coli SHuffle produjo aproximadamente tres veces más cantidades de L1(C30 cuando creció en condiciones de autoinducción, con respecto a la inducción convencional y, por ende, las cantidades fueron comparables a las obtenidas por E. coli BL21(DE3) bajo inducción convencional.  Conclusiones:  La truncación de treinta residuos de aminoácidos por el extremo carboxilo de la proteína L1 del VPH-18 tuvo una importante contribución a la producción y solubilidad de la proteína L1 nativa en E. coli. Este es el primer informe sobre la producción soluble de la proteína L1 del VPH-18 en una cepa de E. coli SHuffle. Sin embargo, se necesitan mayores cantidades de la proteína L1 para escalar su producción para desarrollar un candidato vacunal contra el VPH.]]></p></abstract>
<kwd-group>
<kwd lng="en"><![CDATA[human papillomavirus]]></kwd>
<kwd lng="en"><![CDATA[L1 major capsid protein]]></kwd>
<kwd lng="en"><![CDATA[vaccine, Escherichia coli]]></kwd>
<kwd lng="es"><![CDATA[papiloma humano]]></kwd>
<kwd lng="es"><![CDATA[proteína principal de la cápsida L1]]></kwd>
<kwd lng="es"><![CDATA[vacuna]]></kwd>
<kwd lng="es"><![CDATA[Escherichia coli]]></kwd>
</kwd-group>
</article-meta>
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