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Highly efficient construction of infectious viroid-derived clones

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Highly efficient construction of infectious viroid-derived clones

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dc.contributor.author Márquez-Molins, Joan es_ES
dc.contributor.author NAVARRO BOHIGUES, JOSE ANTONIO es_ES
dc.contributor.author Pallás Benet, Vicente es_ES
dc.contributor.author Gomez, Gustavo Germán es_ES
dc.date.accessioned 2021-01-28T04:31:32Z
dc.date.available 2021-01-28T04:31:32Z
dc.date.issued 2019-08-01 es_ES
dc.identifier.issn 1746-4811 es_ES
dc.identifier.uri http://hdl.handle.net/10251/160075
dc.description.abstract [EN] Background Viroid research generally relies on infectious cDNA clones that consist of dimers of the entire viroid sequence. At present, those dimers are generated by self-ligation of monomeric cDNA, a strategy that presents several disadvantages: (i) low efficiency, (ii) it is a non-oriented reaction requiring tedious screenings and (iii) additional steps are required for cloning into a binary vector for agroinfiltration or for in vitro RNA production. Results We have developed a novel strategy for simultaneous construction of a viroid dimeric cDNA and cloning into a multipurpose binary vector ready for agroinfiltration or in vitro transcription. The assembly is based on IIs restriction enzymes and positive selection and supposes a universal procedure for obtaining infectious clones of a viroid independently of its sequence, with a high efficiency. Thus, infectious clones of one viroid of each family were obtained and its infectivity was analyzed by molecular hybridization. Conclusion This is a zero-background strategy for direct cloning into a binary vector, optimized for the generation of infectious viroids. As a result, this methodology constitutes a powerful tool for viroid research and exemplifies the applicability of type IIs restriction enzymes and the lethal gene ccdB to design efficient and affordable direct cloning approaches of PCR products into binary vectors. es_ES
dc.description.sponsorship This work was supported by the Spanish Ministry of Economy and Competitiveness (co-supported by FEDER) Grants BIO2017-88321-R (VP) and AGL2016-79825-R (GG). The funders had no role in the experiment design, data analysis, decision to publish, or preparation of the manuscript. es_ES
dc.language Inglés es_ES
dc.publisher Springer (Biomed Central Ltd.) es_ES
dc.relation.ispartof Plant Methods es_ES
dc.rights Reconocimiento (by) es_ES
dc.subject Viroid es_ES
dc.subject Cloning es_ES
dc.subject Dimers es_ES
dc.subject IIs enzymes es_ES
dc.subject Agro-infiltration es_ES
dc.title Highly efficient construction of infectious viroid-derived clones es_ES
dc.type Artículo es_ES
dc.identifier.doi 10.1186/s13007-019-0470-4 es_ES
dc.relation.projectID info:eu-repo/grantAgreement/MINECO//AGL2016-79825-R/ES/VALIDACION FUNCIONAL DE LAS REDES DE SNCRNAS QUE REGULAN LA REPUESTA A ESTRES EN MELON. ANALISIS DE SU POTENCIAL COMO FUENTE DE TOLERANCIA A CONDICIONES AMBIENTALES ADVERSAS/ es_ES
dc.relation.projectID info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2017-88321-R/ES/DESCRIFRANDO INTERACCIONES VIRUS-PLANTA ESENCIALES PARA LA SUSCEPTIBILIDAD Y%2FO RESISTENCIA EN DOS PATOSISTEMAS AGRONOMICAMENTE RELEVANTES/ 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.description.bibliographicCitation Márquez-Molins, J.; Navarro Bohigues, JA.; Pallás Benet, V.; Gomez, GG. (2019). Highly efficient construction of infectious viroid-derived clones. Plant Methods. 15:1-8. https://doi.org/10.1186/s13007-019-0470-4 es_ES
dc.description.accrualMethod S es_ES
dc.relation.publisherversion https://doi.org/10.1186/s13007-019-0470-4 es_ES
dc.description.upvformatpinicio 1 es_ES
dc.description.upvformatpfin 8 es_ES
dc.type.version info:eu-repo/semantics/publishedVersion es_ES
dc.description.volume 15 es_ES
dc.identifier.pmid 31388344 es_ES
dc.identifier.pmcid PMC6670230 es_ES
dc.relation.pasarela S\393280 es_ES
dc.contributor.funder European Regional Development Fund es_ES
dc.contributor.funder Agencia Estatal de Investigación es_ES
dc.contributor.funder Ministerio de Economía y Competitividad es_ES
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