Abstract
Carotenoids are widespread pigments in photosynthetic species, but they are also found in nonphotosynthetic microorganisms, such as bacteria and fungi. The amenability of fungi to genetic studies have made some fungal species advantageous models in the study of the genetics and biochemistry of carotenoid biosynthesis, while others have been used for biotechnological carotenoid production. The availability of molecular techniques that allow modulating the expression of target genes is a powerful tool in the manipulation of carotenoid synthesis. An example of an adjustable gene expression is based on the tetracycline-controlled transcriptional activation system, known as Tet-on. We describe here the material and protocols for the construction of a Tet-on regulated gene, its introduction in the filamentous fungus F. fujikuroi, and its use to modulate the expression of a negative regulator of carotenoid biosynthesis.
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Acknowledgments
We thank the Spanish Government (projects BIO2012-39716, BIO2015-69613-R, and CaRed Networks BIO2015-71703-REDT and BIO2017-90877-REDT), and Junta de Andalucía (project CTS-6638) for financial support. J.P-M was granted by Ministry of Science, Innovation and University and P.O had a Fellowship for Initiation to Research from Plan Propio from University of Seville.
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Marente, J., Ortega, P., Pardo-Medina, J., Avalos, J., Limón, M.C. (2020). Modulation of Activity of a Carotenoid Pathway Through the Use of the TeT-on Regulatory System: Application in the Fungus Fusarium fujikuroi. In: Rodríguez-Concepción, M., Welsch, R. (eds) Plant and Food Carotenoids. Methods in Molecular Biology, vol 2083. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9952-1_26
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DOI: https://doi.org/10.1007/978-1-4939-9952-1_26
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