Obtención y caracterización parcial de un mutante de la proteína recombinante α-sinucleína, y producción de su anticuerpo policlonal

dc.contributorInstituto de Genética de la Universidad Nacional de Colombia – IGUN
dc.contributorRey Buitrago, Mauricio
dc.creatorGantiva Gantiva, Mauricio
dc.date.accessioned2020-02-26T15:44:22Z
dc.date.available2020-02-26T15:44:22Z
dc.date.created2020-02-26T15:44:22Z
dc.date.issued2019
dc.identifierhttps://repositorio.unal.edu.co/handle/unal/75758
dc.description.abstractLa -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.languagespa
dc.publisherUniversidad Nacional de Colombia - Sede Bogotá
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dc.titleObtención y caracterización parcial de un mutante de la proteína recombinante α-sinucleína, y producción de su anticuerpo policlonal
dc.typeOtro


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