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dc.contributor.advisorHernández Rojas, Edith del Carmen
dc.contributor.advisorOrtiz Suárez, Heidy Daniela
dc.contributor.authorAcosta Muñoz, Laura Michell
dc.contributor.authorArias García, Lauren Nayarinne
dc.date.accessioned2021-05-25T18:24:25Z
dc.date.available2021-05-25T18:24:25Z
dc.date.issued2020-03-20
dc.identifier.urihttps://repositorio.unicolmayor.edu.co/handle/unicolmayor/92
dc.description.abstractLa malaria causada por la especie Plasmodium vivax, es una enfermedad de alto impacto en salud pública alrededor del mundo, se reporta que el 53% de los casos de malaria por esta especie se concentran en el Sudeste Asiático, el 47% en la India y en países de América latina representa una amenaza permanente, siendo la especie predominante con alrededor del 75% de los casos de paludismo. P. vivax ha demostrado un interés de investigación debido a su importancia epidemiológica, a pesar de esto los estudios respecto a las interacciones moleculares en el proceso de invasión se han visto retrasados por su difícil propagación in vitro debido a la preferencia del parásito por invadir reticulocitos, como consecuencia de lo anterior el conocimiento sobre las moléculas envueltas en el proceso de invasión, es escaso. En este estudio se expresaron y analizaron estructuralmente dos fragmentos de la proteína PvSPATR, el fragmento 1 con un tamaño 333 de pb y el fragmento 2 con un tamaño de 429 pb a través del uso de técnicas moleculares, como PCR y, técnicas inmersas en la obtención de las proteínas recombinantes. Esta proteína ha sido reportada en P. falciparum como importante para la unión del esporozoito a los hepatocitos y, como proteína multiestadío, lo que sugiere continuar con ensayos de unión y creación de péptidos específicos que determinen la importancia de este antígeno a nivel funcional in vitro y permita así una aproximación de su función in vivo.spa
dc.description.tableofcontentsRESUMEN 1 1. Introducción 2 2. Objetivos 4 3. ANTECEDENTES 5 4. MARCO TEÓRICO 7 4.1 La malaria 7 4.2 Estado actual de la malaria 7 4.2.1 En el mundo 8 4.2.2 En Colombia 9 4.3 Vectores que transmiten la malaria 9 4.4 Generalidades y descripción de Plasmodium spp 9 4.4.1 Morfología y Ciclo de vida 9 4.4.2 Especies de Plasmodium sp. que causan malaria 11 4.4.3 Plasmodium falciparum 11 4.4.4 Plasmodium vivax 11 4.5 Proteínas descritas en Plasmodium implicadas en el reconocimiento e invasión del hepatocito por el esporozoíto 14 4.5.1 Proteínas descritas en P. falciparum 14 4.5.2 Proteínas descritas en P. vivax 15 4.6 Proteína SPATR 15 5.1 Tipo de investigación 17 5.2 Enfoque de la investigación 17 5.3 Población 17 5.4 Muestra 17 5.5 Procedimientos 18 5.5.1 Obtención de la muestra 18 5.5.2 Identificación del gen spatr de P. vivax 18 5.5.3 Diseño de cebadores 18 5.5.4 Amplificación de SPATR por PCR 20 5.5.5 Purificación de producto de PCR 21 5.5.6 PCR mix para adición de adeninas 21 5.5.7 Ligación al vector 22 5.5.8 Transformación 22 5.5.9 Obtención de recombinantes 23 5.5.10 PCR confirmación del inserto 23 5.5.11 Transformación en células BL21-AI para expresión de la proteína 24 5.5.12 Obtención de los fragmentos expresados 24 5.5.12.1 Obtención de forma denaturante 24 5.5.12.2 Obtención de los fragmentos de forma nativa 25 5.5.13 Purificación por cromatografía de afinidad 25 5.5.14 Renaturación de las recombinantes 25 5.5.15 SDS PAGE y Western blot 26 5.5.16 Diálisis 27 5.5.17 Cuantificación de proteínas 27 5.5.17.1 Cuantificación por método BCA 28 5.5.17.2 Cuantificación por método Bradford 29 5.5.18 Dicroísmo Circular 30 6. RESULTADOS 32 6.1 Amplificación de los fragmentos de PvSPATR 32 6.2 Purificación 32 6.3 Obtención de recombinantes 33 6.4 Secuencia de los fragmentos obtenidos 34 6.5 Expresión de PvSPATR recombinante 35 6.6 Purificación de proteína 35 6.7 Cuantificación de proteína 37 6.8 Dicroísmo Circular 37 6.8.1 SPATR Fragmento 1 Renaturalizado 38 6.8.2 Fragmento 2 renaturalizado 39 6.8.3 Fragmento 2 nativo 40 7. DISCUSIÓN 42 8. CONCLUSIONES 46 REFERENCIAS BIBLIOGRÁFICAS 48spa
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dc.format.mimetypeapplication/pdfspa
dc.language.isospaspa
dc.publisherUniversidad Colegio Mayor de Cundinamarcaspa
dc.rightsDerechos Reservados-Universidad Colegio Mayor de Cundinamarca, 2020spa
dc.rights.urihttps://creativecommons.org/licenses/by-nc-sa/4.0/spa
dc.titleClonación y expresión de dos fragmentos de la proteína Spatr de plasmodium vivax en el sistema Escherichia colispa
dc.typeTrabajo de grado - Pregradospa
dc.description.degreelevelPregradospa
dc.description.degreenameBacteriólogo(a) y Laboratorista Clínicospa
dc.publisher.facultyFacultad de Ciencias de la Saludspa
dc.publisher.placeBogota D.C.spa
dc.publisher.programBacteriología y Laboratorio Clínicospa
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dc.rights.accessrightsinfo:eu-repo/semantics/closedAccessspa
dc.rights.creativecommonsAtribución-NoComercial-CompartirIgual 4.0 Internacional (CC BY-NC-SA 4.0)spa
dc.subject.lembcefalea
dc.subject.lembmialgias
dc.subject.lembmosquito Anopheles
dc.subject.proposalMalariaspa
dc.subject.proposalPlasmodium spspa
dc.subject.proposalPlasmodium vivaxspa
dc.subject.proposalproteínas recombinantesspa
dc.subject.proposalPvSPATRspa
dc.type.coarhttp://purl.org/coar/resource_type/c_7a1fspa
dc.type.coarversionhttp://purl.org/coar/version/c_970fb48d4fbd8a85spa
dc.type.contentTextspa
dc.type.driverinfo:eu-repo/semantics/bachelorThesisspa
dc.type.redcolhttps://purl.org/redcol/resource_type/TPspa
dc.type.versioninfo:eu-repo/semantics/publishedVersionspa
dc.rights.coarhttp://purl.org/coar/access_right/c_14cbspa


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