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Obtención y caracterización parcial de un mutante de la proteína recombinante α-sinucleína, y producción de su anticuerpo policlonal
| dc.rights.license | Atribución-NoComercial-SinDerivadas 4.0 Internacional |
| dc.contributor.advisor | Instituto de Genética de la Universidad Nacional de Colombia – IGUN |
| dc.contributor.advisor | Rey Buitrago, Mauricio |
| dc.contributor.author | Gantiva Gantiva, Mauricio |
| dc.date.accessioned | 2020-02-26T15:44:22Z |
| dc.date.available | 2020-02-26T15:44:22Z |
| dc.date.issued | 2019 |
| dc.identifier.uri | https://repositorio.unal.edu.co/handle/unal/75758 |
| dc.description.abstract | La -sinucleína es el componente principal de los cuerpos de Lewy y un sello patogénico de todas las sinucleinopatías, en personas con alto consumo de alcohol, se han observado cambios importantes en la expresión de α sinucleína, alterando la neuroprotección. Las mutaciones en el gen -sinucleína y su correspondiente producción de proteína mutada han sido asociadas con procesos de agregación de la proteína, en donde se ve alterada su conformación y adquiriere capacidad autoagregante, lo que se relaciona con el depósito de agregados proteináceos en las neuronas y podría constituir un factor fisiopatológico importante en enfermedades neuronales. Dentro de las mutaciones identificadas, una mutación importante "G51D" en el gen de la -sinucleína sugirio que la -sinucleína podría servir como marcador anatomopatológico para enfermedades neuronales, y de adicciones relacionadas con el abuso de alcohol. En este proyecto presentamos la metodología que nos permitió establecer las mejores condiciones de expresión, purificación y caracterización de la proteína recombinante α-sinucleína G51D, con ensayos de agragación y la estandarizaron de las condiciones para la obtención del anticuerpo de tipo policlonal dirigido hacia el antígeno α-sinucleína G51D; para lograr lo anterior se identifico por medio de estudios bioinformáticos un candidato mutante de la proteína α-sinucleína, que en nuestro caso resultó en un cambio en el aminoácido 51 de la proteína, siendo reemplazada la Glicina por Acido aspártico; se realizó la optimización para la clonación del gen de estudio, utilizando el vector pET30a, que nos dio la mejor solubilidad in silico, y la proteína obtenida permitió la obtención de un anticuerpo policlonal anti proteína mutada α-sinucleína G51D, constituyendo una herramienta inmunológica importante en la confirmación de la existencia de proteínas mutantes a nivel de la α-sinucleína. |
| dc.format.extent | 88 |
| dc.format.mimetype | application/pdf |
| dc.language.iso | spa |
| dc.rights | Derechos reservados - Universidad Nacional de Colombia |
| dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0/ |
| dc.subject.ddc | Medicina y salud |
| dc.title | Obtención y caracterización parcial de un mutante de la proteína recombinante α-sinucleína, y producción de su anticuerpo policlonal |
| dc.type | Otro |
| dc.rights.spa | Acceso abierto |
| dc.description.additional | Magíster en Genética Humana. Línea de investigación: Genética Clínica |
| dc.type.driver | info:eu-repo/semantics/other |
| dc.type.version | info:eu-repo/semantics/acceptedVersion |
| dc.description.degreelevel | Maestría |
| dc.publisher.branch | Universidad Nacional de Colombia - Sede Bogotá |
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| dc.rights.accessrights | info:eu-repo/semantics/openAccess |
| dc.subject.proposal | α-sinucleína |
| dc.subject.proposal | α-sinucleína G51D |
| dc.subject.proposal | pET30a |
| dc.subject.proposal | Mutante |
| dc.subject.proposal | Anticuerpo policlonal |
| dc.type.coar | http://purl.org/coar/resource_type/c_1843 |
| dc.type.coarversion | http://purl.org/coar/version/c_ab4af688f83e57aa |
| dc.type.content | Text |
| oaire.accessrights | http://purl.org/coar/access_right/c_abf2 |
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