The aim of this study was to evaluate the MPK1 (SLT2) gene deletion upon filamentous growth induced by isoamyl alcohol (IAA) in two haploid industrial strains of Saccharomyces cerevisiae using oligonucleotides especially designed for a laboratory S. cerevisiae strain. The gene deletion was performed by replacing part of the open reading frames from the target gene with the KanMX gene. The recombinant strains were selected by their resistance to G418, and after deletion confirmation by polymerase chain reaction, they were cultivated in a yeast extract peptone dextrose medium + 0.5% IAA to evaluate the filamentous growth in comparison to wild strains. Mpk1 derivatives were obtained for both industrial yeasts showing the feasibility of the oligonucleotides especially designed for a laboratory strain (Σ1278b) by Martinez-Anaya et al. (In yeast, the pseudohyphal phenotype induced by isoamyl alcohol results from the operation of the morphogenesis checkpoint. J Cell Sci 116:3423–3431, 2003). The filamentation rate in these derivatives was significantly lower for both strains, as induced by IAA. This drastic reduction in the filamentation ability in the deleted strains suggests that the gene MPK1 is required for IAA-induced filamentation response. The growth curves of wild and derivative strains did not differ substantially. It is not known yet whether the switch to filamentous growth affects the fermentative characteristics of the yeast or other physiological traits. A genetically modified strain for nonfilamentous growth would be useful for these studies, and the gene MPK1 could be a target gene. The feasibility of designed oligonucleotides for this deletion in industrial yeast strains is shown.
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The authors wish to thank Dr. Peter E. Sudbery (University of Sheffield, UK) for his helpful suggestions during the postdoc supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (no. 200678/01-9) for the second author and Dr. Claudia Martinez-Anaya for providing the oligonucleotides. The research was supported by Fundação de Amparo a Pesquisa do Estado de São Paulo (04/06889-0 and 04/06669-0 grants).
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Vancetto, G.T., Ceccato-Antonini, S.R. MPK1 gene is required for filamentous growth induced by isoamyl alcohol in Saccharomyces cerevisiae strains from the alcoholic fermentation. Appl Microbiol Biotechnol 75, 111–115 (2007). https://doi.org/10.1007/s00253-006-0795-6
- Saccharomyces cerevisiae
- Isoamyl alcohol