Print Email Facebook Twitter amdSYM, a new dominant recyclable marker cassette for Saccharomyces cerevisiae Title amdSYM, a new dominant recyclable marker cassette for Saccharomyces cerevisiae Author Solis-Escalante, D. Kuijpers, N.G.A. Bongaerts, N. Bolat, I. Bosman, L. Pronk, J.T. Daran, J.M. Daran-Lapujade, P.A.S. Faculty Applied Sciences Department BT/Biotechnology Date 2012-12-17 Abstract Despite the large collection of selectable marker genes available for Saccharomyces cerevisiae, marker availability can still present a hurdle when dozens of genetic manipulations are required. Recyclable markers, counterselectable cassettes that can be removed from the targeted genome after use, are therefore valuable assets in ambitious metabolic engineering programs. In the present work, the new recyclable dominant marker cassette amdSYM, formed by the Ashbya gossypii TEF2 promoter and terminator and a codon-optimized acetamidase gene (Aspergillus nidulans amdS), is presented. The amdSYM cassette confers S. cerevisiae the ability to use acetamide as sole nitrogen source. Direct repeats flanking the amdS gene allow for its efficient recombinative excision. As previously demonstrated in filamentous fungi, loss of the amdS marker cassette from S. cerevisiae can be rapidly selected for by growth in the presence of fluoroacetamide. The amdSYM cassette can be used in different genetic backgrounds and represents the first counterselectable dominant marker gene cassette for use in S. cerevisiae. Furthermore, using astute cassette design, amdSYM excision can be performed without leaving a scar or heterologous sequences in the targeted genome. The present work therefore demonstrates that amdSYM is a useful addition to the genetic engineering toolbox for Saccharomyces laboratory, wild, and industrial strains. Subject counter-selectable markerdominant markerscarless marker removalacetamidaseserial gene deletionSaccharomyces cerevisiae To reference this document use: http://resolver.tudelft.nl/uuid:21f3c07b-19ff-48f6-af54-31f7eca13e1e DOI https://doi.org/10.1111/1567-1364.12024 Publisher Blackwell Publishing ISSN 1567-1356 Source FEMS Yeast Research, 13 (1), 2012 Part of collection Institutional Repository Document type journal article Rights © 2012 Federation of European Microbiological Societies Files PDF Solis-Escalante_2012.pdf 1.26 MB Close viewer /islandora/object/uuid:21f3c07b-19ff-48f6-af54-31f7eca13e1e/datastream/OBJ/view