Abstract
Undegraded Vicia faba polysomes from meristematic root cells were obtained after homogenization in a medium of low ionic strength provided that the pH was equal to 9.0. By minimizing the shearing forces during the homogenization step, polysomes were obtained free of mitochondrial and nuclear contaminants, measured by differential spectrophotometry and CsCl gradient centrifugation respectively. Poly(A)-containing RNA was obtained by poly(U)-Sepharose chromatography and shown to be virtually free of rRNA and its average size was 13–15 S. Approximately 9% of the purified preparation was annealed by [3H]-poly(U). Sucrose gradient analysis under denaturing conditions showed that the poly(A)-containing RNA were non-degraded. This RNA was used to direct the synthesis of proteins in a heterologous cell-free system from wheat germ.
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Ryffel, G. U. and McCarthy, B. J., 1975. Biochem. 14, 1379–1385.
Galan, G. A., Klein, W. M., Wold, B. J., Britten, R. J. and Davidson, E. H., 1976. Cell 7, 487–505.
Axel, R., Feigelson, P. and Schutz, G., 1976. Cell 7, 247–254.
Hastie, N. D. and Bishop, J. O., 1976. Cell 9, 761–774.
Young, B. D., Birnie, G. D. and Paul, J., 1976. Biochem. 15, 2823–2829.
Affara, N., Jacquet, M., Jakob, H., Jacob, F. and Gros, F., 1977. Cell 12, 509–520.
Felsani, A., Berthelot, F. and Croizat, B., 1978. Eur. J. Biochem. 92, 569–577.
Sphor, G., Grandboulan, N., Morel, C. and Scherrer, K., 1970. Eur. J. Biochem. 73, 296–318.
Trapy, G. and Esnault, R., 1978. Phytochem. 17, 1859–1861.
Lowry, O. H., Rosebrough, W. J. and Randall, R. J., 1951. J. Biol. Chem. 193, 265–267.
Gilkens, A. L. J., Berns, J. J. M. and Bloemendal, H., 1971. Eur. J. Biochem. 22, 478–484.
Breen, M. D., Whitehead, E. T. and Kenefick, D. G., 1972. Plant Physiol. 49, 733–739.
Beopoulos, N., Esnault, R. and Buri, J. F., 1978. Biochim. Biophys. Acta 517, 216–227.
Roberts, B. E. and Paterson, B. M., 1973. Proc. Natl. Acad. Sci. USA 70, 2330–2334.
Larkins, B. A. and Davies, E., 1975. Plant Physiol. 55, 749–756.
Van de Walle, C., 1973. F.E.B.S. Letters 34, 31–34.
Jackson, A. O. and Larkins, B. A., 1976. Plant Physiol. 57, 5–10.
Weeks, D. P. and Marcus, A., 1969. Plant Physiol. 44, 1291–1294.
Lonsdale, D. M. and Boulter, D., 1973. Phytochem. 12, 39–41.
Sala-Trepat, J. M., Savage, M. J. and Bonner, J., 1978. Biochim. Biophys. Acta 519, 173–193.
Shuster, L., 1957. J. Biol. Chem. 229, 289–303.
Trapy, G., Esnault, R. and Zuily-Fodil, Y., 1975. Plant Sc. Letters 4, 257–265.
Trebal, J. P., Beopoulos, N. and Esnault, R., 1979. Phytochem. 18, 1635–1637.
Dissous, C., Verwaerde, C., Lempereur, C. and Krembel, J., 1978. Eur. J. Biochem. 83, 5–15.
Higgins, T. J. V., Mercer, J. B. and Goodwin, P. B., 1973. Nature New Biol. 246, 68–70.
Delseny, M., Aspart, L., Balay, H. and Guitton, Y., 1974. C.R. Acad. Sci. Paris 278, 1225–1228.
Esnault, R., Trapy, G. and van Huystee, R. B., 1975. Eur. J. Biochem. 60, 83–89.
He, D. T. H. and Varner, J. E., 1974. 71, 4783–4786.
Jacobsen, D. V. and Zwar, J. H., 1974. Proc. Natl. Acad. Sci. USA 71, 3290–3293.
Harris, B. and Dure, L. S., 1974. Biochem. 13, 5463–5467.
Grierson, D. and Covey, S., 1975. Planta 127, 77–86.
Tobin, E. M. and Klein, A. O., 1975. Plant Physiol. 56, 88–92.
Covey, S. and Grierson, D., 1976. Eur. J. Biochem. 63, 599–606.
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Tessier, L., Esnault, R. Isolation and characterization of polysomes and polyadenylated polysomal RNA from vicia faba meristematic root cells. Mol Cell Biochem 29, 173–181 (1980). https://doi.org/10.1007/BF00420288
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DOI: https://doi.org/10.1007/BF00420288