Overexpression And Purification Of Pwl2d, A Mutant Of The Effector Protein Pwl2 From Magnaporthe Grisea.

dc.creatorSchneider, D R S
dc.creatorSaraiva, A M
dc.creatorAzzoni, A R
dc.creatorMiranda, H R C A N
dc.creatorde Toledo, M A S
dc.creatorPelloso, A C
dc.creatorSouza, A P
dc.date2010-Nov
dc.date2015-11-27T13:17:49Z
dc.date2015-11-27T13:17:49Z
dc.date.accessioned2018-03-29T01:10:49Z
dc.date.available2018-03-29T01:10:49Z
dc.identifierProtein Expression And Purification. v. 74, n. 1, p. 24-31, 2010-Nov.
dc.identifier1096-0279
dc.identifier10.1016/j.pep.2010.04.020
dc.identifierhttp://www.ncbi.nlm.nih.gov/pubmed/20438845
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/198833
dc.identifier20438845
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1299066
dc.descriptionThe rice blast disease caused by the ascomycete Magnaporthe grisea continues to cause a tremendous impact in rice (Oryza sativa) cultures around the world. Elucidating the molecular basis of the fungus interactions with its host might help increase the general understanding of the pathogen-host relationship. At the moment of invasion, the fungus secretes effectors that modify host defenses and cellular processes as they successively invade living rice cells. PWL2, an effector protein, is a known AVR (avirulence) gene product. The PWL2 gene prevents the fungus from infecting weeping lovegrass (Eragrostis curvula). In this study, we identified a PWL2 allele gene (which we termed PWL2D) in a strain of M. grisea. The sequence of PWL2D has only two bases different from that of PWL2, producing alterations in residue 90 and residue 142. However, the alteration of residue 90 (from D(90) to N(90)) is critical to gene function. Here, we cloned the gene PWL2D in a pET System vector, expressed the gene product in Escherichia coli and evaluated by spectroscopic techniques some aspects of the PWL2D structure. While TRX-tagged PWL2D is prone to aggregation, the solubility of PWL2D is improved when it is overexpressed without its original signal peptide. Expression and purification procedures for these constructs are described. Finally, we found out that the protein seems to be an intrinsically disordered protein. Results from these studies will facilitate structural analysis of PWL2D and might contribute to understanding the gene's function and of fungal/plant interactions.
dc.description74
dc.description24-31
dc.languageeng
dc.relationProtein Expression And Purification
dc.relationProtein Expr. Purif.
dc.rightsfechado
dc.rightsCopyright 2010 Elsevier Inc. All rights reserved.
dc.sourcePubMed
dc.subjectAlleles
dc.subjectAmino Acid Sequence
dc.subjectCircular Dichroism
dc.subjectCloning, Molecular
dc.subjectEscherichia Coli
dc.subjectFungal Proteins
dc.subjectGenes, Fungal
dc.subjectGenetic Vectors
dc.subjectMagnaporthe
dc.subjectMolecular Sequence Data
dc.subjectMutant Proteins
dc.subjectMutation
dc.subjectNuclear Magnetic Resonance, Biomolecular
dc.subjectProtein Conformation
dc.subjectSequence Alignment
dc.subjectThioredoxins
dc.subjectUp-regulation
dc.titleOverexpression And Purification Of Pwl2d, A Mutant Of The Effector Protein Pwl2 From Magnaporthe Grisea.
dc.typeArtículos de revistas


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