Exploiting multiSite Gateway and penNFRUIT plasmid collection for fruit genetic engineering

[EN] MultiSite Gateway cloning techniques based on homologous recombination facilitate the combinatorial assembly of basic genetic pieces (i.e., promoters, CDS, and terminators) into gene expression or gene silencing cassettes. pENFRUIT is a collection of MultiSite Triple Gateway Entry vectors dedic...

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Detalles Bibliográficos
Autores: Hueso Estornell, Leandro, GRANELL RICHART, ANTONIO|||0000-0003-4266-9581, Orzáez Calatayud, Diego Vicente
Tipo de recurso: artículo
Fecha de publicación:2012
País:España
Institución:Universitat Politècnica de València (UPV)
Repositorio:RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia
Idioma:inglés
OAI Identifier:oai:riunet.upv.es:10251/83432
Acceso en línea:https://riunet.upv.es/handle/10251/83432
Access Level:acceso abierto
Palabra clave:Gateway cloning
Gene expression
Gene silencing
HpRNAi
Transient expression
Agroinjection
BIOQUIMICA Y BIOLOGIA MOLECULAR
Descripción
Sumario:[EN] MultiSite Gateway cloning techniques based on homologous recombination facilitate the combinatorial assembly of basic genetic pieces (i.e., promoters, CDS, and terminators) into gene expression or gene silencing cassettes. pENFRUIT is a collection of MultiSite Triple Gateway Entry vectors dedicated to genetic engineering in fruits. It comprises a number of fruit-operating promoters as well as C-terminal tags adapted to the Gateway standard. In this way, flanking regulatory/labeling sequences can be easily Gateway-assembled with a given gene of interest for its ectopic expression or silencing in fruits. The resulting gene constructs can be analyzed in stable transgenic plants or in transient expression assays, the latter allowing fast testing of the increasing number of combinations arising from MultiSite methodology. A detailed description of the use of MultiSite cloning methodology for the assembly of pENFRUIT elements is presented.