Green fluorescent protein purification through Immobilized Metal Affinity Chromatografy (IMAC) and its relevance for Biomedical Science students during Biochemistry practical classes at La Trobe University – Australia
This work was performed as an integrated practical of a Biomedical Science undergraduate course of Biochemistry subject, in order to demonstrate used techniques to purify of Green Fluorescent Protein (GFP). To perform the experiments the main methodology applied was the by immobilized metal affinity...
| Autores: | , , |
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| Tipo de recurso: | artículo |
| Estado: | Versión publicada |
| Fecha de publicación: | 2016 |
| País: | Brasil |
| Institución: | Sociedade Brasileira de Bioquímica e Biologia Molecular (SBBq) |
| Repositorio: | Revista de Ensino de Bioquímica |
| Idioma: | inglés |
| OAI Identifier: | oai:ojs.bioquimica.org.br:article/638 |
| Acceso en línea: | http://bioquimica.org.br/revista/ojs/index.php/REB/article/view/638 |
| Access Level: | acceso abierto |
| Palabra clave: | practical class, purification GFP; Purification; IMAC Aulas práticas GFP; Purificação; IMAC |
| Sumario: | This work was performed as an integrated practical of a Biomedical Science undergraduate course of Biochemistry subject, in order to demonstrate used techniques to purify of Green Fluorescent Protein (GFP). To perform the experiments the main methodology applied was the by immobilized metal affinity chromatography (IMAC). The open reading frame for enhanced GFP was sub-cloned into the pQE30 expression vector. The subsequent production of protein tagged N-terminally with hexahistidine, facilitated its purification by IMAC. An approximate 3-fold purification of GFP was achieved. Thus, the students who completed the course gained significant experience related to fundamental techniques in molecular cloning and a sound basis in the principles of recombinant protein expression and purification. |
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