Open AccessamdSYM , a new dominant recyclable marker cassette for S accharomyces cerevisiae
Author(s) -
SolisEscalante Daniel,
Kuijpers Niels G.A.,
Bongaerts Nadine,
Bolat Irina,
Bosman Lizanne,
Pronk Jack T.,
Daran JeanMarc,
DaranLapujade Pascale
Publication year - 2013
Publication title -
fems yeast research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.991
H-Index - 92
eISSN - 1567-1364
pISSN - 1567-1356
DOI - 10.1111/1567-1364.12024
Subject(s) - biology , saccharomyces cerevisiae , expression cassette , gene cassette , selectable marker , genetics , ura3 , aspergillus nidulans , computational biology , genome , gene , genetic screen , marker gene , plasmid , mutant , vector (molecular biology) , recombinant dna , integron
Despite the large collection of selectable marker genes available for S accharomyces 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 amd SYM , formed by the A shbya gossypii TEF 2 promoter and terminator and a codon‐optimized acetamidase gene ( A spergillus nidulans amd S ), is presented. The amd SYM cassette confers S . cerevisiae the ability to use acetamide as sole nitrogen source. Direct repeats flanking the amd S gene allow for its efficient recombinative excision. As previously demonstrated in filamentous fungi, loss of the amd S marker cassette from S . cerevisiae can be rapidly selected for by growth in the presence of fluoroacetamide. The amd SYM 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, amd SYM excision can be performed without leaving a scar or heterologous sequences in the targeted genome. The present work therefore demonstrates that amd SYM is a useful addition to the genetic engineering toolbox for S accharomyces laboratory, wild, and industrial strains.