Establishment of a transient and stable transformation protocol of Nicotiana tabacum mediated by Agrobacterium tumefaciens

 

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Detalles Bibliográficos
Autores: Méndez-Muñoz, Laura, Campos-Delgado, So a, Obando-Coronado, Mariel, Garro-Monge, Giovanni
Formato: artículo original
Estado:Versión publicada
Fecha de Publicación:2019
Descripción:Biotechnological tools such as genetic engineering and molecular biology are very advanced, allowing the development of systems based on the use of plant cells for the production of recombinant proteins, which allows the development of platforms for biopharmaceuticals, enzymes and secondary metabolites production. Transient and stable transformation, allow production of recombinant proteins. In this work a stable transformation of N. tabacum callus from the BY-2 commercial line and a wild one (T1) was performed. In addition, agroinfiltration assays were carried out using two techniques: by syringe and vacuum. In both cases, the transformation was mediated by Agrobacterium tumefaciens LBA4404 which carried the vector pCAMBIA 1303, containing the GFP and GUS marker genes, and a hygromycin resistance gene. The expression of green fluorescent protein was evaluated by fluorescence microscopy. Expression of the hygromycin and β-glucuronidase resistance genes were evaluated by PCR. As a result, high GFP expression and good transformation rates were obtained.
País:RepositorioTEC
Institución:Instituto Tecnológico de Costa Rica
Repositorio:RepositorioTEC
Lenguaje:Español
OAI Identifier:oai:repositoriotec.tec.ac.cr:2238/11852
Acceso en línea:https://revistas.tec.ac.cr/index.php/tec_marcha/article/view/4120
https://hdl.handle.net/2238/11852
Access Level:acceso abierto
Palabra clave:Agroinfiltration
agroinfection
fluorescence
BY-2
pCAMBIA 1303
acetosyringone
hygromycin
β-glucuronidase
Agroinfiltración
agroinfección
fluorescencia
acetosiringona
Higromicina
β-glucoronidasa.