Summary
Expression of the β-galactosidase gene in yeast has served as a screening marker for many purposes. Here it is shown that in two yeasts, Saccharomyces cerevisiae and Schizosaccharomyces pombe, the β-glucuronidase (GUS) gene can be used as an alternative marker. Since the histochemical substrate can not be taken up by yeast cells, direct colony screening of plates was found to be impossible. However, by a replica plating technique, GUS expression became visibly detectable within 10 min when the GUS gene was strongly expressed. The staining method could still be performed for expression at a 100-fold lower level, but incubation times of several hours were needed. Furthermore, specific GUS expression levels of yeast protein extracts could be quantified by a fluorometric assay which is both very simple to perform and highly sensitive. Since the GUS gene can also tolerate large N-terminal fusions, this method should be particularly attractive for studying such diverse problems as transcriptional and translational regulation or subcellular localization in yeast.
References
Bassford PJ, Beckwith J, Berman E, Casadaban M, Guarente L, Saint-Girons I, Sarthy A, Schwartz M, Shuman H, Silhavy T (1978) In: Miller JH, Reznikoff WS (eds) The operon. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 245–258
Bradford MM (1976) Anal Biochem 72:248–254
Guarente L, Ptashne M (1981) Proc Natl Acad Sci USA 78:2199–2202
Hirt H, Murbacher T, Kögl M, Heberle-Bors E (1990) Curr Genet 17:473–479
Ito H, Fukuda Y, Murata K, Kimura A (1983) J Bacteriol 153:163–168
Jefferson RA (1987) Plant Mol Biol Rep 5:387–405
Jefferson RA, Burgess SM, Hirsh D (1986) Proc Natl Acad Sci USA 83:8447–8451
Platt T, Müller-Hill B, Miller JH (1972) In: Miller JH (ed.) Experiments in Molecular Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 352–359
Pobjecky N, Rosenberg GH, Dinter-Gottlieb G, Käufer NF (1990) Mol Gen Genet 220:314–316
Rose M, Casadaban MJ, Botstein D (1981) Proc Natl Acad Sci USA 78:2460–2464
Schmitz UK, Lonsdale DM, Jefferson RA (1990) Curr Genet 17:261–264
Stoeber F (1961) These de Docteur de Sciences, Paris
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Communicated by R. J. Schweyen
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Hirt, H. A novel method for in situ screening of yeast colonies with the β-glucuronidase reporter gene. Curr Genet 20, 437–439 (1991). https://doi.org/10.1007/BF00317075
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DOI: https://doi.org/10.1007/BF00317075