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Cloning and heterologous overexpression of three gap genes encoding different glyceraldehyde-3-phosphate dehydrogenases from the plant pathogenic bacterium Pseudomonas syringae pv. tomato strain DC3000

 

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dc.creator Elkhalfi, Bouchra es
dc.creator Araya, José Miguel es
dc.creator Rodríguez Castro, Jorge es
dc.creator Rey, Manuel es
dc.creator Soukri, Abdelaziz es
dc.creator Serrano Delgado, Aurelio es
dc.date.accessioned 2019-04-02T11:53:26Z
dc.date.available 2019-04-02T11:53:26Z
dc.date.issued 2013
dc.identifier.issn 1046-5928 (impreso) es
dc.identifier.issn 1096-0279 (electrónico) es
dc.identifier.uri https://hdl.handle.net/11441/85064
dc.description.abstract The gammaproteobacterium Pseudomonas syringae pv. tomato DC3000 is the causal agent of bacterial speck, a common disease of tomato. The mode of infection of this pathogen is not well understood, but according to molecular biological, genomic and proteomic data it produces a number of proteins that may promote infection and draw nutrients from the plant. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a major enzyme of carbon metabolism that was reported to be a surface antigen and virulence factor in other pathogenic microorganisms, but its possible role in the infection process of P. syringae has so far not been studied. Whole-genome sequence analyses revealed the occurrence in this phytopathogenic bacterium of three paralogous gap genes encoding distinct GAPDHs, namely two class I enzymes having different molecular mass subunits and one class III bifunctional D-erythrose-4-phosphate dehydrogenase/GAPDH enzyme. By using genome bioinformatics data, as well as alignments of both DNA and deduced protein sequences, the three gap genes of P. syringae were one-step cloned with a His-Tag in pET21a vector using a PCR-based strategy, and its expression optimized in E. coli BL21 to achieve high yield of the heterologous proteins. In accordance with their distinct molecular phylogenies, these bacterial gap genes encode functional GAPDHs of diverse molecular masses and nicotinamide-coenzyme specificities, suggesting specific metabolic and/or cellular roles. es
dc.description.sponsorship Agencia Española de Cooperación Internacional y Desarrollo (MAEC) A1/043076/11 A/030965/10 es
dc.description.sponsorship Junta de Andalucía BIO-261 es
dc.format application/pdf es
dc.language.iso eng es
dc.publisher Academic Press es
dc.rights Attribution-NonCommercial-NoDerivatives 4.0 Internacional *
dc.rights.uri http://creativecommons.org/licenses/by-nc-nd/4.0/ *
dc.subject Pseudomonas syringae pv tomato DC3000 es
dc.subject Glyceraldehyde-3-phosphate dehydrogenase es
dc.subject GAPDH es
dc.subject Paralogous gapgenes es
dc.subject Heterologous protein es
dc.subject Optimized overexpression es
dc.title Cloning and heterologous overexpression of three gap genes encoding different glyceraldehyde-3-phosphate dehydrogenases from the plant pathogenic bacterium Pseudomonas syringae pv. tomato strain DC3000 es
dc.type info:eu-repo/semantics/article es
dc.type.version info:eu-repo/semantics/acceptedVersion es
dc.rights.accessrights info:eu-repo/semantics/openAccess es
dc.contributor.affiliation Universidad de Sevilla. Departamento de Bioquímica Vegetal y Biología Molecular es
dc.relation.projectID A1/043076/11 es
dc.relation.projectID A/030965/10 es
dc.relation.projectID BIO-261 es
dc.relation.publisherversion http://dx.doi.org/10.1016/j.pep.2013.02.005 es
dc.identifier.doi 10.1016/j.pep.2013.02.005 es
idus.format.extent 10 p. es
dc.journaltitle Protein Expression and Purification es
dc.publication.volumen 89 es
dc.publication.initialPage 146 es
dc.publication.endPage 155 es
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