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Chromosomal promoter replacement in Saccharomyces cerevisiae: Construction of conditional lethal strains for the cloning of glycosyltransferases from various organisms


Mazhari-Tabrizi, R; Blank, M; Mumberg, D; Funk, M; Schwarz, RT; Eckert, V
1999-11
GLYCOCONJUGATE JOURNAL
Journal Article
16
11
673-679
Heterologous complementation in yeast has been a successful tool for cloning and characterisation of genes from various organisms. Therefore we constructed conditionally lethal Saccharomyces cerevisiae strains by replacing the endogenous promoter from the genes of interest (glycosyltransferases) by the stringently regulated GAL1-promoter, by a technique called chromosomal promoter replacement. Such yeast strains were constructed for the genes Alg 1, Alg7, Sec59, Wbp1 involved in N-Glycosylation, the genes Gpi2, Gpi3/Spt14, Gaal, Pis1, involved in GPI-anchor biosynthesis and Dpm involved in both pathways. All strains show the expected conditionally lethal phenotype on glucose-containing medium when expression of the respective gene is turned off.
KLUWER ACADEMIC PUBL
PHOSPHATE MANNOSE SYNTHASE; GPI-ANCHOR; GENE DELETION; YEAST; EXPRESSION; PROTEIN; PHOSPHATIDYLINOSITOL; GLYCOSYLATION; BIOSYNTHESIS; DISRUPTION
10.1023/A:1007132907235
Refer to copyright notice on published article.

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Creation Date 1999-11-01 12:00:00