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A micropreparative chromatographic strategy for the purification and sequence analysis of murine granulocyte-colony stimulating factor (mG-CSF)

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Summary

A micropreparative chromatographic strategy using short (<5 cm) 1 and 2 mm ID HPLC columns for preparing pure mG-CSF and its peptide fragments is described. The proteins and peptides are used to obtain both N-terminal and internal amino acid sequence data for the purpose of constructing sequence specific oligonucleotide probes for use in the isolation of the gene coding for the protein of interest, thereby ultimately enabling amplified amounts of protein to be generated using prokaryotic expression systems.

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References

  1. A. W. Burgess, N. A. Nicola, in “Growth Factors and Stem Cells”, Academic Press, Sydney, 1983.

    Google Scholar 

  2. T. Maniatis, E. F. Fritsch, J. Sembrook, in “Molecular Cloning”, Cold Spring Harbor Press, New York, 1982.

    Google Scholar 

  3. T. Huynh, R. A. Young, R. W. Davis, in “DNA cloning: A Practical Approach”, D. M. Glover, ed., IRL Press, Oxford.

  4. R. M. Hewick, M. W. Hunkapiller, L. E. Hood, W. J. Dreyer, J. Biol. Chem.256, 7990 (1981).

    CAS  Google Scholar 

  5. M. W. Hunkapiller, L. E. Hood, Science219, 650 (1983).

    Article  CAS  Google Scholar 

  6. N. A. Nicola, D. Metcalf, M. Matsumoto, G. R. Johnson, J. Biol. Chem.258, 9017 (1983).

    CAS  Google Scholar 

  7. J. M. Bower, R. Camble, H. Gregory, E. L. Gerring, I. R. Willshire, Experientia (Basel)31, 825 (1975).

    CAS  Google Scholar 

  8. B. Grego, I. R. Van Driel, P. A. Stearne, J. W. Goding, E. C. Nice, R. J. Simpson, Eur. J. Biochem.148, 485 (1985).

    Article  CAS  Google Scholar 

  9. E. C. Nice, M. Capp, M. J. O’Hare, J. Chromatogr.185, 413 (1979).

    Article  CAS  Google Scholar 

  10. E. C. Nice, C. J. Lloyd, A. W. Burgess, J. Chromatogr.29, 153 (1984).

    Article  Google Scholar 

  11. E. C. Nice, B. Grego, R. J. Simpson, Biochem. Int.11, 87 (1985).

    Google Scholar 

  12. M. Kunitani, P. Hirtzer, D. Johnson, R. Halenbeck, A. Boosman, K. Koths, J. Chromatogr.359, 391 (1986).

    Article  CAS  Google Scholar 

  13. E. C. Nice, L. Fabri, L. Sparrow, In preparation.

  14. N. A. Nicola, D. Metcalf, J. Cell Physiol.124, 313 (1985).

    Article  CAS  Google Scholar 

  15. B. S. Welinder, S. Linde, J. S. Brush, J. Chromatogr.257, 162 (1983).

    Article  CAS  Google Scholar 

  16. R. J. Simpson, J. A. Smith, R. L. Moritz, M. J. O’Hare, P. S. Rudland, J. R. Morrison, C. J. Lloyd, B. Grego, A. W. Burgess, E. C. Nice, Eur. J. Biochem.153, 629 (1985).

    Article  CAS  Google Scholar 

  17. S. Nagata, M. Tsuchiya, S. Asano et al., Nature,319, 415 (1986).

    Article  CAS  Google Scholar 

  18. R. J. Simpson, N. Nicola, E. C. Nice, Submitted to “Protein Sequences and Data Analysis”.

  19. F. S. Esch, Anal. Biochem.136, 39 (1984).

    Article  CAS  Google Scholar 

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Nice, E.C., Simpson, R.J. & Nicola, N.A. A micropreparative chromatographic strategy for the purification and sequence analysis of murine granulocyte-colony stimulating factor (mG-CSF). Chromatographia 24, 449–454 (1987). https://doi.org/10.1007/BF02688525

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  • DOI: https://doi.org/10.1007/BF02688525

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