Use este identificador para citar ou linkar para este item: https://locus.ufv.br//handle/123456789/23270
Tipo: Artigo
Título: Molecular characterization and expression profile of pectin-lyase-encoding genes from Penicillium griseoroseum
Autor(es): Bazzolli, Denise S.
Ribon, Andrea O. B.
Queiroz, Marisa V. de
Araujo, Elza F. de
Abstract: Penicillium griseoroseum has been studied by our group because of its good pectinase production. Attempts have been done to clone pectinolytic genes, aiming to obtain pectinase-overproducing strains for industrial purposes. Here, two genes coding for pectin lyase were isolated from the P. griseoroseum genome. The plg1 gene has an open reading frame of 1341 bp coding for a putative protein of 374 amino acids with a calculated molecular mass of 40.1 kDa. The plg2 gene is characterized by an open reading frame of 1400 nucleotides and codes for a polypeptide of 383 amino acids. The plg1 gene 5'-flanking region contains putative binding sites for the transcription factors involved in regulation by ambient pH and catabolite repression. The primary structure of Plg1 and Plg2 proteins showed a relatively high homology (varying between 32.4% and 74.8%) to fungal pectin lyases characterized to date. Southern blotting analysis revealed that both genes are present as single copies in the fungus genome. Expression studies revealed a differing pattern of gene expression of plg1 and plg2 when mycelium was cultivated on medium containing different pectic components. Citric pectin followed by apple pectin were the carbon sources that best induced plg1 expression, and transcripts were detected from 24 to 76 h. The expression of the plg2 gene was monitored by reverse transcriptase--polymerase chain reaction, since Northern analysis failed to detect hybridization signals. The differential expression of these genes may provide means for the fungus to adapt to various growth conditions.
Penicillium griseoroseum a ete etudie par notre groupe a cause de sa bonne production de pectinase. Nous avons tente de cloner les genes pectolytiques dans le but d'obtenir des souches surproduisant de la pectinase pour un usage industriel. Dans le cas present, deux genes codant pour des pectine lyases furent isoles du genome de P. griseoroseum. Le gene plg1 a un cadre de lecture ouvert de 1341 paires de base codant une proteine putative de 374 acides amines ayant un masse moleculaire calcule de 40,1 kDa. Le gene plg2 est caracterise par un cadre de lecture ouvert de 1400 nucleotides et code un polypeptide de 383 acides amines. La region 5' flanquant le gene plg1 contient des sites de liaison putatifs pour des facteurs de transcription impliques dans la regulation par le pH ambiant et la repression catabolique. La structure primaire des proteines Plg1 et Plg2 demontrait une homologie relativement elevee (variant entre 32,4 % et 74,8 %) a des pectine lyases fongiques caracterisees jusqu'a maintenant. Une analyse d'hybridation de type Southern a revele que les deux genes etaient presents en tant que copies simples dans le genome du champignon. Des etudes d'expression ont revele un patron d'expression genique qui differait entre plg1 et plg2 lorsque le mycelium etait cultive dans un milieu contenant differentes composantes pectiques. La pectine citrique, suivie par la pectine de pommes, etait la source de carbone qui induisait le mieux l'expression de plg1 et ses transcrits furent detectes de 24 a 76 h. L'expression du gene plg2 fut relevee par reaction de la polymerase en chaine et transcription inverse puisque l'analyse par Northern n'a pu detecter de signal d'hybridation. L'expression differentielle de ces genes pourrait fournir au champignon le moyen de s'adapter a differentes conditions de croissance.
Palavras-chave: Pectin lyase
Gene cloning
Penicillium griseoroseum
Gene expression.
Pectine lyase
Clonage de genes
Penicillium griseoroseum
Expression genique
Editor: Canadian Journal of Microbiology
Tipo de Acesso: NRC Canada
URI: https://doi.org/10.1139/w06-070
http://www.locus.ufv.br/handle/123456789/23270
Data do documento: 2006
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