Author/Editor     Hansen, Jorgen; Felding, Troels; Johannesen, Pia Francke; Piškur, Jure; Christensen, Christina L; Olesen, Kjeld
Title     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
Type     članek
Source     FEMS Yeast Res
Vol. and No.     Letnik 4, št. 3
Publication year     2003
Volume     str. 323-7
Language     eng
Abstract     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.
Descriptors     GENES, REPORTER
LAC OPERON
ACETYLTRANSFERASES
FUNGAL PROTEINS
GENETIC MARKERS
GENETIC VECTORS
HEXOSYLTRANSFERASES
PHOSPHOTRANSFERASES (ALCOHOL GROUP ACCEPTOR)
PLASMIDS
REPLICATION ORIGIN
SACCHAROMYCES CEREVISIAE