Abstract
While carrying out a systematic disruption of the genes of unknown function in the alc gene cluster from the filamentous fungus Aspergillus nidulans, we observed a strong diminution of the transcription of markers inserted in the alcS gene. This was found to be the case for the two markers tested, nadA (from A. nidulans) and pyrG (from A. fumigatus) involved in purine utilization and uracil/uridine biosynthetic pathway, respectively. The same phenomenon was also observed with insertion of the nadA gene in the alcM locus, another gene of the alc cluster. In the case of nadA, the level of expression was directly correlated to the ability of the corresponding strains to grow on adenine as a sole nitrogen source. The insertion of the pyrG marker into alcS complemented perfectly vegetative growth, but did not allow a proper sexual cycle. This suggests that the lowered pyrG expression is not sufficient to provide the intracellular concentration of pyrimidines required for the sexual cycle. Thus, due caution must be exercised when disrupting genes with pyrG, one of the most commonly employed markers, especially if the gene to be disrupted is involved or suspected to be involved in the sexual cycle.
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Acknowledgments
This work was supported by the Centre National de la Recherche Scientifique (CNRS: contract ATIP 2JE077). X.R. was a recipient of a doctoral fellowship from the Ministère de l’Enseignement Supérieur et de la Recherche of the French Government and was then supported by the Human Earth Foundation. We thank Andrew Pearson for correcting the English. The co-authors are grateful to Dr. Michel Flipphi and Prof. Claudio Scazzocchio for fruitful discussions and valuable advice.
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Communicated by G. Braus.
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Robellet, X., Oestreicher, N., Guitton, A. et al. Gene silencing of transgenes inserted in the Aspergillus nidulans alcM and/or alcS loci. Curr Genet 56, 341–348 (2010). https://doi.org/10.1007/s00294-010-0303-5
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DOI: https://doi.org/10.1007/s00294-010-0303-5