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Multigene Engineering by GoldenBraid Cloning: From Plants to Filamentous Fungi and Beyond

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Multigene Engineering by GoldenBraid Cloning: From Plants to Filamentous Fungi and Beyond

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dc.contributor.author Vázquez-Vilar, Marta es_ES
dc.contributor.author Gandía, Mónica es_ES
dc.contributor.author García-Carpintero, Victor es_ES
dc.contributor.author Marqués, Eric es_ES
dc.contributor.author Sarrion-Perdigones, Alejandro es_ES
dc.contributor.author Yenush, Lynne es_ES
dc.contributor.author Polaina, Julio es_ES
dc.contributor.author Manzanares, Paloma es_ES
dc.contributor.author Marcos, Jose F. es_ES
dc.contributor.author Orzáez Calatayud, Diego Vicente es_ES
dc.date.accessioned 2021-05-14T03:31:41Z
dc.date.available 2021-05-14T03:31:41Z
dc.date.issued 2020-03-09 es_ES
dc.identifier.issn 1934-3639 es_ES
dc.identifier.uri http://hdl.handle.net/10251/166341
dc.description This is the peer reviewed version of the following article: Vazquez-Vilar, M., Gandía, M., García-Carpintero, V., Marqués, E., Sarrion-Perdigones, A., Yenush, L., Polaina, J., Manzanares, P., Marcos, J. F., & Orzaez, D. (2020). Multigene engineering by goldenbraid cloning: from plants to filamentous fungi and beyond. Current Protocols in Molecular Biology, 130, e116, doi: 10.1002/cpmb.116, which has been published in final form at https://doi.org/10.1002/cpmb.116. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving. es_ES
dc.description.abstract [EN] Many synthetic biologists have adopted methods based on Type IIS restriction enzymes and Golden Gate technology in their cloning procedures, as these enable the combinatorial assembly of modular elements in a very efficient way following standard rules. GoldenBraid (GB) is a Golden Gate¿based modular cloning system that, in addition, facilitates the engineering of large multigene constructs and the exchange of DNA parts as result of its iterative cloning scheme. GB was initially developed specifically for plant synthetic biology, and it has been subsequently extended and adapted to other organisms such as Saccharomyces cerevisiae, filamentous fungi, and human cells by incorporating a number of host¿specific features into its basic scheme. Here we describe the general GB cloning procedure and provide detailed protocols for its adaptation to filamentous fungi¿a GB variant known as FungalBraid. The assembly of a cassette for gene disruption by homologous recombination, a fungal¿specific extension of the GB utility, is also shown. Development of FungalBraid was relatively straightforward, as both plants and fungi can be engineered using the same binary plasmids via Agrobacterium¿mediated transformation. We also describe the use of a set of web¿based tools available at the GB website that assist users in all cloning procedures. The availability of plant and fungal versions of GB will facilitate genetic engineering in these industrially relevant organisms. es_ES
dc.description.sponsorship This article is dedicated to the memory of our friend and colleague Dr. Alejandro Sarrion-Perdigones, an early developer of GoldenBraid. We acknowledge the excellent technical assistance provided by Marisol Gascón (IBMCP, Valencia, Spain) with the fluorescent images. This work was funded by Grant BIO2013- 42193 and Grant BIO2016-78601-R, Plan Nacional I+D, Spanish Ministry of Economy and Competitiveness, RTI2018-101115-B-C21 from the Ministerio de Ciencia, Innovación y Universidades (Spain) (MICINN/FEDER Funds), and PROMETEO/ 2018/066 from Conselleria d'Educació (Generalitat Valenciana, Comunitat Valenciana, Spain) and SUSPHIRE PCI2018- 092893-ERA CoBioTech (109) (MCIU/FEDER). es_ES
dc.language Inglés es_ES
dc.publisher Wiley es_ES
dc.relation.ispartof Current Protocols in Molecular Biology es_ES
dc.rights Reserva de todos los derechos es_ES
dc.subject Agrobacterium-mediated transformation es_ES
dc.subject FungalBraid es_ES
dc.subject Golden-Braid es_ES
dc.subject Modular cloning es_ES
dc.subject Synthetic biology es_ES
dc.subject.classification MICROBIOLOGIA es_ES
dc.subject.classification BIOQUIMICA Y BIOLOGIA MOLECULAR es_ES
dc.title Multigene Engineering by GoldenBraid Cloning: From Plants to Filamentous Fungi and Beyond es_ES
dc.type Artículo es_ES
dc.identifier.doi 10.1002/cpmb.116 es_ES
dc.relation.projectID info:eu-repo/grantAgreement/MINECO//BIO2013-42193-R/ES/GREEN SWITCHES: DISEÑO DE CIRCUITOS GENETICOS ARTIFICIALES PARA LA PRODUCCION DE PROTEINAS RECOMBINANTES Y EL ENRIQUECIMIENTO NUTRICIONAL DE PLANTAS SOLANACEAS/ es_ES
dc.relation.projectID info:eu-repo/grantAgreement/MINECO//BIO2016-78601-R/ES/DISEÑO DE CIRCUITOS GENICOS SINTETICOS Y ORTOGONALES PARA PLANTAS MEDIANTE EL USO DE FACTORES PROGRAMABLES DE UNION A DNA BASADOS EN LA ARQUITECTURA CRISPR-CAS9./ es_ES
dc.relation.projectID info:eu-repo/grantAgreement/AEI//PCI2018-092893/ES/BIOPRODUCCION SOSTENIBLE DE FEROMONAS DE INSECTOS PARA CONTROL DE PLAGAS/ es_ES
dc.relation.projectID info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-101115-B-C21/ES/PROTEINAS ANTIFUNGICAS DE HONGOS: DESDE LOS HONGOS A LAS PLANTAS Y MAS ALLA/ es_ES
dc.relation.projectID info:eu-repo/grantAgreement/GVA//PROMETEO%2F2018%2F066/ es_ES
dc.rights.accessRights Abierto es_ES
dc.contributor.affiliation Universitat Politècnica de València. Instituto Universitario Mixto de Biología Molecular y Celular de Plantas - Institut Universitari Mixt de Biologia Molecular i Cel·lular de Plantes es_ES
dc.contributor.affiliation Universitat Politècnica de València. Departamento de Biotecnología - Departament de Biotecnologia es_ES
dc.description.bibliographicCitation Vázquez-Vilar, M.; Gandía, M.; García-Carpintero, V.; Marqués, E.; Sarrion-Perdigones, A.; Yenush, L.; Polaina, J.... (2020). Multigene Engineering by GoldenBraid Cloning: From Plants to Filamentous Fungi and Beyond. Current Protocols in Molecular Biology. 130(1):1-31. https://doi.org/10.1002/cpmb.116 es_ES
dc.description.accrualMethod S es_ES
dc.relation.publisherversion https://doi.org/10.1002/cpmb.116 es_ES
dc.description.upvformatpinicio 1 es_ES
dc.description.upvformatpfin 31 es_ES
dc.type.version info:eu-repo/semantics/publishedVersion es_ES
dc.description.volume 130 es_ES
dc.description.issue 1 es_ES
dc.identifier.pmid 32150346 es_ES
dc.relation.pasarela S\427152 es_ES
dc.contributor.funder Generalitat Valenciana es_ES
dc.contributor.funder European Regional Development Fund es_ES
dc.contributor.funder Ministerio de Economía y Competitividad es_ES
dc.contributor.funder Agencia Estatal de Investigación es_ES
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dc.subject.ods 02.- Poner fin al hambre, conseguir la seguridad alimentaria y una mejor nutrición, y promover la agricultura sostenible es_ES


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