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| Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL | DATACITE | |
| Campo DC | Valor | Lengua/Idioma |
|---|---|---|
| dc.contributor.author | Tallada, Víctor A. | - |
| dc.contributor.author | Simanis, Viesturs | - |
| dc.contributor.author | Hagan, Iain M. | - |
| dc.date.accessioned | 2016-02-23T11:51:31Z | - |
| dc.date.available | 2016-02-23T11:51:31Z | - |
| dc.date.issued | 2014 | - |
| dc.identifier | doi: 10.1371/journal.pone.0097683 | - |
| dc.identifier | issn: 1932-6203 | - |
| dc.identifier.citation | PLoS ONE 9(5): e97683 (2014) | - |
| dc.identifier.uri | http://hdl.handle.net/10261/129347 | - |
| dc.description | This is an open-access article distributed under the terms of the Creative Commons Attribution License.-- et al. | - |
| dc.description.abstract | Targeted alteration of the genome lies at the heart of the exploitation of S. pombe as a model system. The rate of analysis is often determined by the efficiency with which a target locus can be manipulated. For most loci this is not a problem, however for some loci, such as fin1+, rates of gene targeting below 5% can limit the scope and scale of manipulations that are feasible within a reasonable time frame. We now describe a simple modification of transformation procedure for directing integration of genomic sequences that leads to a 5-fold increase in the transformation efficiency when antibiotic based dominant selection markers are used. We also show that removal of the pku70+ and pku80+ genes, which encode DNA end binding proteins required for the non-homologous end joining DNA repair pathway, increases the efficiency of gene targeting at fin1+ to around 75-80% (a 16-fold increase). We describe how a natMX6/rpl42+ cassette can be used for positive and negative selection for integration at a targeted locus. To facilitate the evaluation of the impact of a series of mutations on the function of a gene of interest we have generated three vector series that rely upon different selectable markers to direct the expression of tagged/untagged molecules from distinct genomic integration sites. pINTL and pINTK vectors use ura4+ selection to direct disruptive integration of leu1+ and lys1+ respectively, while pINTH vectors exploit nourseothricin resistance to detect the targeted disruption of a hygromycin B resistance conferring hphMX6 cassette that has been integrated on chromosome III. Finally, we have generated a series of multi-copy expression vectors that use resistance to nourseothricin or kanamycin/G418 to select for propagation in prototrophic hosts. Collectively these protocol modifications and vectors extend the versatility of this key model system. | - |
| dc.description.sponsorship | Swiss National Science Foundation (www.snf.ch). École polytechnique fédérale de Lausanne (www.epfl.ch). Cancer Research UK [CRUK] C147/A6058. | - |
| dc.publisher | Public Library of Science | - |
| dc.relation.isversionof | Publisher's version | - |
| dc.rights | openAccess | - |
| dc.title | Extending the Schizosaccharomyces pombe molecular genetic toolbox | - |
| dc.type | artículo | - |
| dc.identifier.doi | 10.1371/journal.pone.0097683 | - |
| dc.relation.publisherversion | http://dx.doi.org/10.1371/journal.pone.0097683 | - |
| dc.date.updated | 2016-02-23T11:51:31Z | - |
| dc.description.version | Peer Reviewed | - |
| dc.language.rfc3066 | eng | - |
| dc.rights.license | http://creativecommons.org/licenses/by/4.0/ | - |
| dc.contributor.funder | Cancer Research UK | - |
| dc.contributor.funder | Swiss National Science Foundation | - |
| dc.contributor.funder | École Polytechnique Fédérale de Lausanne | - |
| dc.relation.csic | Sí | - |
| dc.identifier.funder | http://dx.doi.org/10.13039/501100000289 | es_ES |
| dc.identifier.funder | http://dx.doi.org/10.13039/501100001703 | es_ES |
| dc.identifier.pmid | 24848109 | - |
| dc.type.coar | http://purl.org/coar/resource_type/c_6501 | es_ES |
| item.fulltext | With Fulltext | - |
| item.openairecristype | http://purl.org/coar/resource_type/c_18cf | - |
| item.openairetype | artículo | - |
| item.cerifentitytype | Publications | - |
| item.grantfulltext | open | - |
| Aparece en las colecciones: | (CABD) Artículos | |
Ficheros en este ítem:
| Fichero | Descripción | Tamaño | Formato | |
|---|---|---|---|---|
| molecular genetic toolbox.pdf | 1,87 MB | Adobe PDF |  Visualizar/Abrir |
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