Abstract
By using the renaturation kinetics technique we tried to get informations about the maintenance of the 2 μm plasmid in yeast cells. For this purpose we determined the 2 μm plasmid copy number: in various yeast strains, in a special set of mutants, in cells treated with ethidium bromide and cycloheximide and in different yeast strains obtained by transformation with 2 μm chimeric plasmids.
According to the strain used the proportion of 2μm DNA varied from 1.1% to 3.9%, which corresponds to 24 to 88 2 μm molecules per haploid genome. The particular multiresistant mutant, where the frequent loss of oligomycine resistance is correlated with the loss of extractible covalently closed circular DNA, contained 39 2 μm copies per haploid genome. In the partial revertant oligomycine sensitive all the 2 μm DNA sequences were lost. (Less than 0.1 copy per haploid genome.)
Ethidium bromide did not affect the 2 μm copy number while cycloheximide induces an increase of 36%.
When a strain containing 88 2 μm DNA copies per haploid genome is transformed with 2 μm chimeric plasmids there is no significative change in the total number of plasmid: 36 copies of endogenous and 44 of chimeric plasmid per haploid genome. When 2 μm chimeric plasmids were introduced in our 2 μm-less strain despite the stability of the transformants, there is only 8 copies per haploid genome.
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Bach, M. L., Lacroute, F., Botstein, D.: Proc. Nat. Acad. Sci. USA 76, 386–390 (1979)
Beggs, J. D.: Nature 73, 104–109 (1978)
Blanc, H., Gerbaud, C., Slonimski, P. P., Guerineau, M.: Mol. Gen. Genet. 176, 335–342 (1979)
Bouanchaud, D. H., Scavizzi, M. R., Chabbert, Y. A.: J. Gen. Microbiol. 54, 417–425 (1969)
Bukhari, A. J., Shapiro, J. A., Adhya, S. L. (eds.): DNA insertion elements, plasmids and episomes. Cold Spring Harbor, N. Y.: Cold Spring Harbor Laboratory 1977
Burton, K.: Biochem. J. 62, 315–323 (1956)
Clewell, D. B.: J. Bacteriol. 110, 667–676 (1972)
Clark-Walker, G. D.: Proc. Nat. Acad. Sci. USA 69, 388–392 (1972)
Clark-Walker, G. D., Miklos, G.L.G.: Eur. J. Biochem. 41, 359–365 (1974)
Colbere-Garapin, F., Chousterman, S., Horodniceanu, F., Kourilski, P., Garapin, A. C.: Proc. Nat. Acad. Sci. USA 76, 3755–3759 (1979)
Fournier, P., Gerbaud, C., Blanc, H., Aigle, M., Heslot, H., Guerineau, M.: Use of spheroplasts to transform yeast with chimeric plasmids. In: Proceedings of the 5th international protoplast symposium. Oxford: Pergamon Press and Budapest: Akademiai Kiadi (in press 1980)
Gelb, L. D., Kohne, D. E., Martin, M. A.: J. Mol. Biol. 57, 129–145 (1971).
Gerbaud, C., Fournier, P., Blanc, H., Aigle, M., Heslot, H., Guerineau, M.: Gene 5, 233–253 (1979)
Griffiths, D. E., Lancashire, W. E., Zanders, E. D.: FEBS Lett. 53, 126–130 (1975)
Grossman, L., I., Goldring, E., S., Marmur, J.: J. Mol. Biol. 46, 367–376 (1969)
Guérineau, M., Slonimski, P. P., Avner, P. R.: Biochem. Biophys. Res. Commun. 61, 462–469 (1974)
Guérineau, M., Grandchamp, C., Slonimski, P. P.: Structure and genetic of the 2 μm circular DNA in yeast, in: Genetics and biogenesis of chloroplasts and mitochondria. Th. Bücher et al. (eds), pp. 557–564. Amsterdam: Elsevier/North-Holland - Biomedical Press 1976.
Guérineau, M.: FEBS Lett. 80, 426–428 (1977)
Guérineau, M.: Plasmid DNA in yeast. In: Viruses and Plasmids in fungi. Lemke, P., A. (ed.), pp 539–593. New York: Marcel Dekker 1979
Gubbins, E. J., Newton, C. S., Kann, M. D., Donelson J. E.: Gene 1, 185–207 (1977)
Guerry, J. P., Le Blanc, D. J., Falkow, S.: J. Bacteriol. 116, 1064–1066 (1973)
Hollenberg, C. P., Degelmann, A., Kustermann Kuhn, B., Royer, H. D.: Proc. Nat. Acad. Sci. USA 73, 2072–2076 (1976)
Lauer, G. D., Roberts, T. M., Klotz, L. L.: J. Mol. Biol. 114, 507–526 (1977)
Livingston, D. M., Kupfer, D. M.: J. Mol. Biol. 116, 249–260 (1977).
Mounoulou, J. C., Jakob, H., Slonimski, P. P.: Biochem. Biophys. Res. Com. 24, 218–224 (1966)
Netter, P., Petrochilo, E., Slonimski, P. P., Bolotin-Fukuhara, M., Coen, D., Deutsh, J., Dujon, B.: Genetics 78, 1063–1100 (1974)
Rubin, G. M.: Preparation of RNA and ribosomes from yeast. In: Methods in cell biology. Prescott, D. M. (eds), Vol. XII. pp. 45–64. New York: Academic Press 1975
Rubin, S. J., Rosemblum, E. D.: J. Bacteriol. 108, 1200–1204 (1971).
Saunders, G. W., Rank, G. H., Kustermann Kuhn, B., Hollenberg, C. P.: Mol. Gen. Genet. 175, 45–52 (1979).
Sharp, P. A., Petterson, V., Sambrook, J.: J. Mol. Biol. 86, 709–726 (1974)
Slonimski, P. P., Perrodin, G., Croft, J. H.: Biochem. Biophys. Res. Com. 30, 232–239 (1968)
Struhl, K., Stinchomb, D. T., Scherrer, S., Davis, R. W.: Proc. Nat. Acad. Sci. USA 75, 1035–1039 (1979)
Thuring, R. W. J., Sanders, J. P. M., Borst, P.: Anal. Biochem. 66, 213–220 (1975)
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Gerbaud, C., Guérineau, M. 2 μm plasmid copy number in different yeast strains and repartition of endogenous and 2 μm chimeric plasmids in transformed strains. Curr Genet 1, 219–228 (1980). https://doi.org/10.1007/BF00390947
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DOI: https://doi.org/10.1007/BF00390947