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Cell-free Protein Synthesis resulting in Active Phage Qβ Replicase

Abstract

SEVERAL steps in the replication of the small phage Qβ can be reconstructed under cell-free conditions. Net synthesis of biologically active RNA “plus” strands has been achieved in vitro using RNA from virus particles as a template and RNA dependent RNA polymerase or “replicase” from infected E. coli cells1. This reaction proceeded through complementary RNA or “minus” strands as intermediate templates2. The plus strand-dependent reaction required a “host factor” HF I in addition to purified replicase3–6,7. Furthermore, three of four phage coded proteins can be synthesized in vitro using Qβ RNA as an mRNA and an extract from uninfected E. coli as a protein synthesizing system8,9. Coat protein and the phage specific subunit of replicase (R protein) when synthesized in vitro became bound to Qβ RNA8,10 as would be expected for the authentic proteins. Cell-free synthesis of enzymatically active Qβ replicase, however, has not been reported so far.

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References

  1. Spiegelman, S., Haruna, I., Pace, N. R., Mills, D. R., Bishop, D. H. L., Claybrook, J. R., and Peterson, R., J. Cell Physiol, 70, Suppl. 1, 35 (1967).

    Article  CAS  Google Scholar 

  2. Feix, G., Pollet, R., and Weissmann, C., Proc. natn. Acad. Sci. U.S.A., 59, 145 (1968).

    Article  CAS  Google Scholar 

  3. Shapiro, L., France de Fernandez, M. T., and August, J. T., Nature, 220, 478, (1968).

    Article  CAS  Google Scholar 

  4. Kamen, R., Kondo, M., Römer, W., and Weissmann, C., Eur. J. Biochem., 31, 44 (1972).

    Article  CAS  Google Scholar 

  5. Hori, K., Eoyang, L., Banerjee, A. K., and August, J. T., Proc. natn. Acad. Sci. U.S.A., 57, 1790 (1967).

    Article  CAS  Google Scholar 

  6. Franze de Fernandez, M. T., Haywood, W. S., and August, J. T., J. biol. Chem., 247, 824, (1972).

    CAS  PubMed  Google Scholar 

  7. Eikhom, T. S., and Spiegelman, S., Proc. natn. Acad. Sci. U.S.A., 57, 1833 (1967).

    Article  CAS  Google Scholar 

  8. Jockusch, H., Ball, L. A., and Kaesberg, P., Virology, 42, 401 (1970).

    Article  CAS  Google Scholar 

  9. Horiuchi, K., Webster, R., and Matsuhashi, S., Virology, 45, 429 (1971).

    Article  CAS  Google Scholar 

  10. Jockusch, H., and Hindennach, I., Eur. J. Biochem., 29, 353 (1972).

    Article  CAS  Google Scholar 

  11. Kondo, M., Gallerani, R., and Weissmann, C., Nature, 228, 525 (1970).

    Article  CAS  Google Scholar 

  12. Kamen, R., Nature, 228, 527 (1970).

    Article  CAS  Google Scholar 

  13. Blumenthal, T., Landers, T. A., and Weber, K., Proc. natn. Acad. Sci. U.S.A., 69, 1313 (1972).

    Article  CAS  Google Scholar 

  14. Groner, Y., Sheps, R., Kamen, R., Kolakofsky, D., and Revel, M., Nature new Biol., 239, 19 (1972).

    Article  CAS  Google Scholar 

  15. Kondo, M., and Weissmann, C., Eur.J. Biochem., 24, 530 (1972).

    Article  CAS  Google Scholar 

  16. Eoyang, L., and August, J. T., in Methods in Enzymology (edit. by Grossman, L., and Moldave, K.), 530 (Academic Press, New York, 1968).

    Google Scholar 

  17. Kamen, R., Biochim. biophys. Acta, 262, 88 (1972).

    Article  CAS  Google Scholar 

  18. Happe, M., thesis, Univ. Tübingen (1973).

    Google Scholar 

Download references

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HAPPE, M., JOCKUSCH, H. Cell-free Protein Synthesis resulting in Active Phage Qβ Replicase. Nature New Biology 245, 141–143 (1973). https://doi.org/10.1038/newbio245141a0

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