Further development of the cassette-based pYC plasmid system by incorporation of the dominant hph, nat and AUR1-C gene markers and the lacZ reporter system.
Dominant selection markers encoding hygromycin B phosphotransferase (hph), nourseothricin N-acetyltransferase (nat) and a mutant inositol phosphoceramide synthase (AUR1-C) were all incorporated into the pYC yeast plasmid vector system, thus expanding this system with possible alternatives to the use of G418 resistance. We found the markers to be of use not only in standard laboratory strains of Saccharomyces cerevisiae but also in an industrial strain of S. carlsbergensis (syn. of S. pastorianus) brewing yeast as well as in Saccharomyces kluyveri. As the pYC system contains means of counter-selection for plasmid loss and loop-out of integrated plasmids, it now provides ample opportunities for genetic manipulation of industrial and non-conventional yeasts when the URA3 marker and FOA counter-selection is not an option. Furthermore, the lacZ system for analyzing gene expression was included in the system.
Hansen
Jørgen
J
Carlsberg Research Laboratory, Gamle Carlsberg Vej 10, 2500, Copenhagen-Valby, Denmark.
Felding
Troels
T
Johannesen
Pia Francke
PF
Piskur
Jure
J
Christensen
Christina Lund
CL
Olesen
Kjeld
K
D000123:Acetyltransferases / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D017930:Genes, Reporter; D005819:Genetic Markers; D005822:Genetic Vectors; D006602:Hexosyltransferases / Q000235:genetics; D007763:Lac Operon; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D010957:Plasmids / Q000235:genetics*; D018741:Replication Origin / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development
D005656:Fungal Proteins; D005819:Genetic Markers; D000123:Acetyltransferases; C050473:nourseothricin acetyltransferase; D006602:Hexosyltransferases; C088714:phosphatidylinositol-ceramide phosphoinositol transferase; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase