<?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>2215-3470</journal-id>
<journal-title><![CDATA[Uniciencia]]></journal-title>
<abbrev-journal-title><![CDATA[Uniciencia]]></abbrev-journal-title>
<issn>2215-3470</issn>
<publisher>
<publisher-name><![CDATA[Universidad Nacional, Costa Rica]]></publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id>S2215-34702024000100288</article-id>
<article-id pub-id-type="doi">10.15359/ru.38-1.16</article-id>
<title-group>
<article-title xml:lang="es"><![CDATA[Clonación y caracterización del gen codificante de la proteína de choque térmico HSP83 de Trypanosoma cruzi]]></article-title>
<article-title xml:lang="en"><![CDATA[Cloning and Characterization of the Gene Encoding the Heat Shock Protein HSP83 from Trypanosoma cruzi]]></article-title>
<article-title xml:lang="pt"><![CDATA[Clonagem e caracterização do gene que codifica a proteína de choque térmico HSP83 do Trypanosoma cruzi]]></article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Fernández]]></surname>
<given-names><![CDATA[Ana]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[De Lima]]></surname>
<given-names><![CDATA[Ana Rita]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname><![CDATA[Ferrer]]></surname>
<given-names><![CDATA[Elizabeth]]></given-names>
</name>
<xref ref-type="aff" rid="Aff"/>
</contrib>
</contrib-group>
<aff id="Af1">
<institution><![CDATA[,Universidad de Carabobo Instituto BioMolP, Facultad de Ciencias de la Salud Laboratorio de Protozoología]]></institution>
<addr-line><![CDATA[Valencia ]]></addr-line>
<country>Venezuela</country>
</aff>
<aff id="Af2">
<institution><![CDATA[,Universidad de Carabobo Facultad de Ciencias de la Salud Sede Aragua Instituto de Investigaciones Biomédicas &#8220;Dr. Francisco J. Triana Alonso&#8221; (BIOMED)]]></institution>
<addr-line><![CDATA[Maracay estado Aragua]]></addr-line>
<country>Venezuela</country>
</aff>
<aff id="Af3">
<institution><![CDATA[,Universidad de Carabobo Facultad de Ciencias de la Salud Sede Aragua Instituto de Investigaciones Biomédicas &#8220;Dr. Francisco J. Triana Alonso&#8221; (BIOMED)]]></institution>
<addr-line><![CDATA[Maracay estado Aragua]]></addr-line>
<country>Venezuela</country>
</aff>
<pub-date pub-type="pub">
<day>00</day>
<month>12</month>
<year>2024</year>
</pub-date>
<pub-date pub-type="epub">
<day>00</day>
<month>12</month>
<year>2024</year>
</pub-date>
<volume>38</volume>
<numero>1</numero>
<fpage>288</fpage>
<lpage>302</lpage>
<copyright-statement/>
<copyright-year/>
<self-uri xlink:href="http://www.scielo.sa.cr/scielo.php?script=sci_arttext&amp;pid=S2215-34702024000100288&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.sa.cr/scielo.php?script=sci_abstract&amp;pid=S2215-34702024000100288&amp;lng=en&amp;nrm=iso"></self-uri><self-uri xlink:href="http://www.scielo.sa.cr/scielo.php?script=sci_pdf&amp;pid=S2215-34702024000100288&amp;lng=en&amp;nrm=iso"></self-uri><abstract abstract-type="short" xml:lang="es"><p><![CDATA[Resumen  (Objetivo)  Tryapanosoma cruzi, un agente causal de la enfermedad de Chagas, es un parásito que presenta una alternancia en su ciclo de vida entre un hospedador invertebrado (triatomino) y uno vertebrado (mamífero). Varios estudios han demostrado que en T. cruzi, HSP83 (homólogo de HSP90) es esencial para la división celular y el control de la respuesta al estrés térmico. Esta investigación se concentró en el estudio de la clonación, la caracterización bioinformática y la expresión del gen de la proteína de choque térmico HSP83 de T. cruzi para estudios posteriores sobre señalización.  (Metodología)  El ARN fue extraído de epimagostigotes T. cruci (clon EPm6/MOHM/VE/2007/ 6c), utilizando un kit comercial. El ADNc que codifica HSP83 se obtuvo utilizando RT-PCR, a partir del ARNm extraído, para lo cual se diseñaron los cebadores con base en la secuencia HSP83 de la cepa T. cruzi CL Brener. La clonación se realizó usando pGEM®T-Easy y se subclonó en el vector de expresión pQE30. Se efectuó la caracterización bioinformática y de secuencias. El gen se expresó y la proteína recombinante se purificó por cromatografía de afinidad, igual que se identificó por inmunotransferencia.  (Resultados) El análisis de secuencia mostró similitud con el gen HSP83 de Trypanosoma cruzi y se observaron dominios HSP, así como epítopos B en la secuencia. Después de 3 horas de inducción con IPTG, se obtuvo una proteína recombinante con un peso aproximado de 83 kDa. La reacción de inmunotransferencia con suero hiperinmune anti-epimastigote de T. cruzi permitió la detección de una sola banda con un peso molecular de aproximadamente 83 kDa.  (Conclusiones)  Todos los resultados indican que se logró la clonación y caracterización de HSP83 de Trypanosoma cruzi.]]></p></abstract>
<abstract abstract-type="short" xml:lang="en"><p><![CDATA[Abstract  (Objective)  Trypanosoma cruzi, a causal agent of Chagas&#8217; disease, is a parasite whose life cycle alternates between an invertebrate (triatomine) and a vertebrate (mammal) host. Various studies have shown that in T. cruzi, HSP83 (a homolog of HSP90) is essential for cell division and control of the response to thermal stress. This investigation focused on studying the cloning, bioinformatic characterization, and expression of the heat shock protein HSP83 T. cruzi gene for further cell signaling studies.  (Methodology)  RNA was extracted from T. cruzi epimastigotes (EPm6 clone, MHOM/VE/2007/ 6c) using a commercial kit. The cDNA encoding HSP83 was determined using RT-PCR on extracted mRNA, for which the primers were designed based on the HSP83 sequence of T. cruzi strain CL Brener. Cloning was performed using pGEM®T-Easy and subcloned into the expression vector pQE30. Sequence and bioinformatic characterization were performed. The gene was expressed, and the recombinant protein was purified using affinity chromatography and identified through immunoblotting.  (Results)  Sequence analysis showed similarity to the gene encoding HSP83 from Trypanosoma cruzi, and HSP domains and B epitopes in the sequence were also observed. After 3 hours of induction with IPTG, a recombinant protein with an approximate weight of 83 kDa was obtained. The immunoblotting reaction with hyperimmune anti-T. cruzi epimastigote serum helped detect a single band with a molecular weight of nearly 83 kDa.  (Conclusions)  All results indicate that the cloning and characterization of HSP83 from Trypanosoma cruzi was achieved.]]></p></abstract>
<abstract abstract-type="short" xml:lang="pt"><p><![CDATA[Resumo  (Objetivo)  O Tryapanosoma cruzi, agente causador da doença de Chagas, é um parasita que apresenta uma alternância em seu ciclo de vida entre um hospedeiro invertebrado triatomíneo) e um vertebrado(mamífero). Vários estudos demonstraram que, no T. cruzi, a HSP83 (homóloga da HSP90) é essencial para a divisão celular e o controle da resposta ao estresse térmico. Esta pesquisa concentrou-se no estudo da clonagem, caracterização bioinformática e expressão do gene da proteína de choque térmico HSP83 do T. cruzi para estudos adicionais de sinalização.  (Metodologia)  O RNA foi extraído de epimastigotas de T. cruzi (clone EPm6/MOHM/VE/2007/ 6c) usando um kit comercial. O cDNA que codifica a HSP83 foi obtido por meio de RT-PCR a partir do mRNA extraído, para o qual os primers foram projetados com base na sequência da HSP83 da cepa T. cruzi CL Brener. A clonagem foi realizada com o pGEM®T-Easy e subclonada no vetor de expressão pQE30. Foi realizada a caracterização bioinformática e de sequência. O gene foi expresso e a proteína recombinante foi purificada por cromatografia de afinidade e identificada por imunotransferência.  (Resultados)  A análise da sequência mostrou similaridade com o gene HSP83 do Trypanosoma cruzi e os domínios HSP, bem como os epítopos B, foram observados na sequência. Após 3 horas de indução com IPTG, foi obtida uma proteína recombinante com peso aproximado de 83 kDa. A imunotransferência com soro hiperimune antiepimastigota de T. cruzi permitiu a detecção de uma única banda com peso molecular de aproximadamente 83 kDa.  (Conclusões)  Todos os resultados indicam que a clonagem e a caracterização da HSP83 do Trypanosoma cruzi foram alcançadas.]]></p></abstract>
<kwd-group>
<kwd lng="en"><![CDATA[Trypanosoma cruzi]]></kwd>
<kwd lng="en"><![CDATA[heat shock protein]]></kwd>
<kwd lng="en"><![CDATA[HSP83]]></kwd>
<kwd lng="en"><![CDATA[cloning]]></kwd>
<kwd lng="en"><![CDATA[expression]]></kwd>
<kwd lng="es"><![CDATA[Trypanosoma cruzi]]></kwd>
<kwd lng="es"><![CDATA[proteína de choque térmico]]></kwd>
<kwd lng="es"><![CDATA[HSP83]]></kwd>
<kwd lng="es"><![CDATA[clonación]]></kwd>
<kwd lng="es"><![CDATA[expresión]]></kwd>
<kwd lng="pt"><![CDATA[Trypanosoma cruzi]]></kwd>
<kwd lng="pt"><![CDATA[proteína de choque térmico]]></kwd>
<kwd lng="pt"><![CDATA[HSP83]]></kwd>
<kwd lng="pt"><![CDATA[clonagem]]></kwd>
<kwd lng="pt"><![CDATA[expressão]]></kwd>
</kwd-group>
</article-meta>
</front><back>
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