Cloning and characterization of the glyceraldehyde-3-phosphate dehydrogenase gene and the use of its promoter for expression in Myrothecium gramineum, a novel expression host

Maeseneire, Sofie L. De; Dauvrin, T.; Jonniaux, Jean-Luc; Bogaert, Inge Van; Soetaert, Wim; Vandamme, Erick

doi:10.1111/j.1574-6968.2008.01091.x

Cited by 7 publications

(7 citation statements)

References 14 publications

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“…In many fungi GAPDH is encoded by a single copy gene Hirano et al 1999;De Maeseneire et al 2008), but some fungi have several slightly different gapdh genes. Saccharomyces cerevisiae and Kluyveromyces marxianus have three gapdh genes, and these gapdh genes are differentially expressed (McAlister and Holland 1985b;Fernandes et al 1995).…”

Section: Discussionmentioning

confidence: 99%

Cloning and expression analysis of putative glyceraldehyde-3- phosphate dehydrogenase genes in Pilobolus crystallinus

Kubo

2011

Mycoscience

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Putative glyceraldehyde-3-phosphate dehydrogenase (gapdh) genes were cloned from Pilobolus crystallinus with degenerated primers designed from conserved sequences in many GAPDHs. P. crystallinus had three gapdh homologue genes, named pcgapdh1, pcgapdh2, and pcgapdh3. Deduced amino acid sequences for PCGAPDH1, PCGAPDH2, and PCGAPDH3 showed highest similarity with GPD3, GPD1, and GPD2, respectively, of Mucor circinelloides, indicating that these three gapdh genes had diverged before Pilobolus and Mucor were separated. The expression patterns of the gapdh genes, however, were quite different between P. crystallinus and M. circinelloides. All the three pcgapdh genes were expressed, and the expression of pcgapdh2 was suppressed by glucose and sodium acetate. These results indicate that the function of these orthologous genes was changed after Pilobolus and Mucor were separated.

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Section: Discussionmentioning

confidence: 99%

Cloning and expression analysis of putative glyceraldehyde-3- phosphate dehydrogenase genes in Pilobolus crystallinus

Kubo

2011

Mycoscience

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“…T rich region). Although in almost all the fungal promoters analyzed only the ct-box is found (Kuo et al 2004;Nitta et al 2004;Vastag et al 2004;Fei et al 2006;De Maeseneire et al 2008), both boxes have been found in few of them (Punt et al 1990;Liao et al 2008). In these scarce cases, different fragment promoters have been used to produce recombinant proteins, concluding that although the proximal ct-box alone seems enough to drive the expression of a protein, the maximal expression levels would be reached in the presence of the additional gpd-box (Punt et al 1990(Punt et al , 1992Liao et al 2008).…”

Section: Introductionmentioning

confidence: 98%

Production of a heterologous recombinant protein using fragments of the glyceraldehyde-3-phosphate dehydrogenase promoter from Penicillium camemberti

Espinosa

Trebotich

Sepúlveda

et al. 2011

World J Microbiol Biotechnol

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The biotechnological applications of cheeseripening fungi have been limited by a lack of genetics tools, in particular the identification and characterization of suitable promoters for protein expression. In this study, the suitability of the glyceraldehyde-3-phosphate dehydrogenase (gpdP) promoter from Penicillium camemberti to drive the production of a recombinant protein was evaluated. The gpdP gene and its promoter were isolated using PCR and Genome Walker. The promoter of gpdP has two regions with high identity to the regulatory elements gpd-box and ct-box previously described in Aspergillus nidulans. Two fragments of the promoter containing the gpd-and ct-box or the ct-box alone were used to drive the in vivo production of recombinant b-galactosidase using A. nidulans as host. Our results indicate that larger fragment containing gpd-box enhances the production of b-galactosidase activity levels respect to ct-box alone, and that both boxes are necessary to obtain maximal enzymatic activity production. The smaller fragment (187 nt) containing the ct-box alone was able to trigger up to 27% of b-galactosidase activity, and to our knowledge this is the smallest fragment from a gpd gene used to produce a recombinant protein. Differences were not observed when glycerol, galactose or glucose were used as carbon sources, suggesting that the promoter activity is carbohydrate-independent. This is the first report in which a Penicillium gpd promoter is used for recombinant protein production. Our results open the way for the future development of a system for recombinant proteins expression in the biotechnologically important cheese-ripening fungus P. camemberti.

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“…Although it is encoded by a single-copy gene in many fungi (Hirano et al 1999;De Maeseneire et al 2008;Van Bogaert et al 2008), some yeasts, mushrooms, and filamentous fungi possess several slightly different gapdh genes. Saccharomyces cerevisiae expresses three gapdh genes (McAlister and Holland 1985), Agaricus bisporus Lange possesses two gapdh genes (gpd1 and gpd2), although transcripts have only been detected for one of these genes (Harmsen et al 1992), and Mucor circinelloides Schipper and Pilobolus crystallinus (Wiggers) Tode each have three gapdh genes (gpd1, gpd2 and gpd3, and pcgapdh1, pcgapdh2 and pcgapdh3, respectively) (Wolff and Arnau 2002;Kubo 2011).…”

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confidence: 99%

“…Although GAPDH is indispensable for all living cells, it is encoded by a single-copy gene in most filamentous fungi (Hirano et al 1999;De Maeseneire et al 2008;Van Bogaert et al 2008). M. circinelloides possesses three gapdh genes (gpd1, gpd2 and gpd3), but only gpd1 is known to be expressed (Wolff and Arnau 2002).…”

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confidence: 99%

Asexual reproductive organ-specific expression of the glyceraldehyde-3-phosphate dehydrogenase 2 gene of Pilobolus crystallinus

Kubo

2012

Mycoscience

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Pilobolus crystallinus has three putative glyceraldehyde-3-phosphate dehydrogenase (gapdh) genes (pcgapdh1, pcgapdh2 and pcgapdh3). The results of this study demonstrate that expression of pcgapdh2 was increased by irradiation and that this increased expression was correlated with the formation of asexual reproductive organs (trophocysts). Interestingly, expression of pcgapdh2 was restricted to trophocysts. The formation of trophocysts was likely promoted by light, and the expression of pcgapdh2 was increased as a result of trophocyst formation. This is the first report that shows the regulation of a gapdh gene in an organ-specific manner in fungi.

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Cloning and characterization of the glyceraldehyde-3-phosphate dehydrogenase gene and the use of its promoter for expression in Myrothecium gramineum, a novel expression host

Cited by 7 publications

References 14 publications

Cloning and expression analysis of putative glyceraldehyde-3- phosphate dehydrogenase genes in Pilobolus crystallinus

Cloning and expression analysis of putative glyceraldehyde-3- phosphate dehydrogenase genes in Pilobolus crystallinus

Production of a heterologous recombinant protein using fragments of the glyceraldehyde-3-phosphate dehydrogenase promoter from Penicillium camemberti

Asexual reproductive organ-specific expression of the glyceraldehyde-3-phosphate dehydrogenase 2 gene of Pilobolus crystallinus

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