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Published on: June 13, 2021
Upstream regulatory regions controlling the expression of the Candida utilis maltase gene
Hana Boňková1, Michaela Osadská1, Ján Krahulec1
1Comenius University in Bratislava, Faculty of Natural Sciences, Department of Molecular Biology, Mlynská dolina, 842 15 Bratislava 4, Slovak Republic.
Abstract:
Candida utilis represents a promising expression host, generating relatively high levels of recombinant proteins. The current study presents preliminary characterization of the upstream regulatory regions controlling the carbon source-dependent expression of the C. utilis maltase gene. Cellobiose and soluble starch were recognised as inducers of maltase promoter, besides maltose. We successfully applied the Cre-loxP system to acquire a null mutant strain with disrupted maltase gene and promoter in polyploid yeast C. utilis. Furthermore, the strength and minimal functional region of the promoter was evaluated by measuring β-galactosidase activity. Our results suggest, the qPCR was shown itself a very smart method for relatively easy characterization of the transformants and correlation of the expression levels with gene dosage.
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