Summary
Transformants of sensitive yeast strains containing an expressed cDNA copy of the yeast killer toxin-immunity gene could be selected for by exposure to added killer toxin. For strain AH-22 the transformation frequency was approximately 10% that obtained by selection for leucine prototrophy. The procedure required time for expression of immunity prior to selection, and a screening step to remove non-transformed survivors. Under conditions where active toxin was produced, transformants containing the toxin-immunity gene were at a selective advantage, and cells losing the plasmid were killed. This resulted in self selection of transformants, and affords a way of maintaining plasmid stability in protrophic strains.
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Bostian KA, Elliot Q, Bussey H, Burn V, Smith A, Tipper DJ (1984) Cell 36:741–751
Brown AJP, Goodey AR, Tubb RS (1981) J Inst Brew (London) 87:234–238
Burrows S (1970) Baker's yeast. In: Rose AH, Harrison JS (eds) The yeasts, 3 Yeast technology. Academic Press, London, pp 349–420
Bussey H, Saville D, Hutchins K, Palfree RGE (1979) J Bacteriol 140:888–892
Harrison JS, Graham JCJ (1970) Yeasts in distillery practice. In: Rose AH, Harrison JS (eds) The yeasts, 3 Yeast technology. Academic Press, London, pp 283–348
Henderson RCA (1983) D-phil thesis. Botany School, Oxford University, England
Hinnen A, Hicks JB, Fink GR (1978) Proc Natl Acad Sci USA 75:1929–1933
Ito H, Fukuda Y, Musata K, Kimura A (1983) J Bacteriol 153:163–168
Kaster KR, Burgett SG, Ingolia TD (1984) Curr Genet 8:353–358
Kosaric N, Ng DCM, Russell I, Stewart GG (1980) Ethanol production by fermentation: an alternative liquid fuel. In: Perlman D (ed) Advances in applied microbiology 26. Academic Press, New York, pp 147–227
Lolle S, Skipper N, Bussey H, Thomas DY (1984) EMBO 3:1383–1387
Luria SE, Delbrück M (1943) Genetics 28:491–511
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular Cloning A Laboratory Manual. Cold Spring Harbor Laboratory, pp 86–91
Maule AP, Thomas PD (1973) J Inst Brew (London) 79:137–141
Southern EM (1975) J Mol Biol 98:503–517
Walmsley RM, Gardner DCJ, Oliver SG (1983) Mol Gen Genet 192:361–365
Webster TD, Dickson RC (1983) Gene 26:243–252
Young TW, Yagiu M (1978) Antonie van heeuwenhoek J Microbiol Serol 44:59–77
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Bussey, H., Meaden, P. Selection and stability of yeast transformants expressing cDNA of an Ml killer toxin-immunity gene. Curr Genet 9, 285–291 (1985). https://doi.org/10.1007/BF00419957
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DOI: https://doi.org/10.1007/BF00419957