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Rapid detection of allelic recombination at the gall locus in yeast by assay of the recombinant gene product

Published online by Cambridge University Press:  14 April 2009

Leland H. Johnston
Affiliation:
Division of Microbiology, National Institute for Medical Research, Mill Hill, London NW7 1AA, England

Summary

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A diploid heteroallelic at the gal1 locus and producing little residual galactokinase activity was chosen. Following either γ- or UV-irradiation, recombination occurred leading to the formation of wild-type GAL1 genes and an increase in detectable galactokinase. An enhanced level of enzyme was first detectable 2 h after irradiation and reached a maximum within 12–14 h. With increasing doses of irradiation, more enzyme was produced and this increase superficially resembled that obtained for plated recombinants. However, far more enzyme was actually synthesized than expected from the number of viable recombinants in the cells assayed. This suggests that recombination must have occurred in cells without colony-forming ability, indeed it may have occurred more frequently in these cells than in viable cells.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1982

References

REFERENCES

Adams, B. G. (1972). Induction of galactokinase in Saccharomyces cerevisiae: kinetics of induction and glucose effects. Journal of Bacteriology 111, 308315.Google Scholar
Banks, G. R., Holloman, W. K., Kairis, M. V., Spanos, A. & Yarranton, G. T. (1976). A DNA polymerase from Ustilago maydis. I. Purification and properties of the polymerase activity. European Journal of Biochemistry 62, 131142.Google Scholar
Birge, E. A. & Low, K. B. (1974). Detection of transcribable recombination products following conjugation in rec +, rec B and rec C strains of E. coli K12. Journal of Molecular Biology 83, 447457.CrossRefGoogle Scholar
Chang, L. M. S. (1977). DNA polymerases from baker's yeast. Journal of Biological Chemistry 252, 18731880.Google Scholar
Douglas, H. C. & Hawthorne, D. C. (1964). Enzymatic expression and genetic linkage of genes controlling galactose utilisation in Saccharomyces. Genetics 49, 837844.CrossRefGoogle ScholarPubMed
Douglas, H. C. & Hawthorne, D. C. (1966). Regulation of genes controlling synthesis of the galactose pathway enzymes in yeast. Genetics 54, 911916.Google Scholar
Game, J. C., Zamb, T. J., Braun, R. J., Resnick, M. & Roth, R. M. (1980). The role of radiation (rad) genes in meiotic recombination in yeast. Genetics 94, 5168.Google Scholar
Holliday, R. (1971). Biochemical measure of the time and frequency of radiation-induced allelic recombination in Ustilago. Nature New Biology 232, 233236.Google Scholar
Joshi, G. P. & Siddiqi, O. (1968). Enzyme synthesis following conjugation and recombination in E. coli. Journal of Molecular Biology 32, 201210.CrossRefGoogle Scholar
Nogi, Y., Matsumoto, K., Toh-e, A. & Oshima, Y. (1977). Interaction of super-suppressible and dominant constitutive mutations for the synthesis of galactose pathway enzymes in Saccharomyces cerevisiae. Molecular and General Genetics 152, 137144.Google Scholar
Parks, J. S., Gottesman, P., Perlman, R. L. & Pastan, I. (1971). Regulation of galactokinase synthesis by cAMP in cell-free extracts. Journal of Biological Chemistry 246, 24192424.CrossRefGoogle Scholar
Prakash, S., Prakash, L., Burke, W. & Montelone, B. A. (1980). Effects of the rodS2 gene on recombination in Saccharomyces cerevisiae. Genetics 94, 3150.Google Scholar
Pritchard, R. H. (1963). The relationship between conjugation, recombination and DNA synthesis in E. coli. In: Genetics Today. Proceedings Eleventh International Congress of Genetics, vol 2 (ed. Geerts, S. J.), pp. 5568. Pergamon Press, Oxford.Google Scholar
Wickerham, L. J. (1951). Taxonomy of yeasts. U.S. Department of Agriculture, Technical Bulletin, no 1029 G.P.O. Washington, DC.Google Scholar
Wintersburger, U. & Blutsch, H. (1976). DNA polymerase from yeast mitochondria. European Journal of Biochemistry 68, 199206.Google Scholar