Abstract
Reporter genes consisting of sporulation-specific promoters fused to lacZ were used as markers to monitor the sporulation pathway of the yeast Saccharomyces cerevisiae. Strains transformed with these lacZ gene fusions expressed β-galactosidase (assayable on plates using the substrate 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside, X-gal) in a sporulation-dependent manner. Mutagenesis experiments performed on transformed strains resulted in the recovery of a number of novel sporulation mutants. Three classes of mutants were obtained: those which overexpressed the reporter gene under sporulation conditions, those which did not express the gene under any conditions, and those which expressed the gene in vegetative cells not undergoing sporulation. On the basis of the blue colony-colour produced in the presence of X-gal these have been described as superblue, white, and blue vegetative mutants, respectively. These were further characterised using earlier reporter genes and other marker systems. This study established that the multicopy reporter plasmids chosen do not interfere with sporulation; they are valid tools for monitoring the pathway and they provide a way to isolate mutations not readily selected by other markers.
References
Briza P, Ellinger A, Winkler G, Breitenbach M (1990) J Biol Chem 265:15118–15123
Casadaban MJ, Martinez-Arias A, Shapira SK, Chou J (1983) Methods Enzymol 100:293–308
Coe JGS, Murray LE, Kennedy CJ, Dawes IW (1992) Mol Microbiol 6:75–81
Dawes IW, Hardie ID (1974) Mol Gen Genet 131:281–289
Esposito MS, Esposito RE (1969) Genetics 61:79–89
Esposito RE, Klapholz S (1981) In: Strathern JW, Jones EW, Broach JR (eds) Molecular biology of the yeast Saccharomyces cerevisiae: life cycle and inheritance. Cold Spring Harbor Laboratory. Cold Spring Harbor, New York, pp 211–287
Feinberg AP, Vogelstein B (1983) Anal Biochem 132:6–13
Grunstein M, Hogness DS (1975) Proc Natl Acad Sci USA 72:3961–3965
Guarente L (1983) Methods Enzymol 101:181–191
Hartwell LH (1974) Bacteriol Rev 38:164–198
Hoffman CS, Winston F (1987) Gene 57:267–272
Holaway BL, Kao G, Finn MC, Clancy MJ (1987) Mol Gen Genet 210:449–459
Ito H, Fukuda Y, Murata K, Kimura A (1983) J Bacteriol 153:163–168
Kao G, Mannix DG, Holaway BL, Finn MC, Bonny AE, Clancy MJ (1989) Mol Gen Genet 215:490–500
Lindegren G, Hwang YL, Oshima Y, Lindegren CC (1965) Can J Genet Cytol 7:491–499
Mortimer RK, Hawthorne DC (1973) Genetics 74:33–54
Ogur M, St John R, nagai S (1957) Science 125:928–929
Roman H, Jacob F (1958) Cold Spring Harbor Symp Quant Biol 23:155–160
Strich R, Slater MR, Esposito RE (1989) Proc Natl Acad Sci USA 86:10018–10022
Williamson DH, Fennell DJ (1975) Methods Cell Biol 12:335–351
Author information
Authors and Affiliations
Additional information
Communicated by M. S. Esposito
Rights and permissions
About this article
Cite this article
Gurvitz, A., Coe, J.G.S. & Dawes, I.W. Use of reporter genes for the isolation and characterisation of different classes of sporulation mutants in the yeast Saccharomyces cerevisiae . Curr Genet 24, 451–454 (1993). https://doi.org/10.1007/BF00351856
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00351856