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Biotechnology Techniques

, Volume 10, Issue 10, pp 737–742 | Cite as

Isolation and partial characterization of the Asp-B10, Tyr-B25-des-(B26-B30)-proinsulin analog from inclusion bodies in Escherichia coli

  • A. M. A. Nascimento
  • H. R. T. Souza
  • M. S. Xavier
  • J. E. Thiemann
  • L. Vilela
  • M. Mares-Guia
Article
  • 18 Downloads

Summary

A proinsulin analog constructed by site-directed mutagenesis was expressed as a fusion protein that formed inclusion bodies inside the cells. It was purified from the isolated inclusion bodies and proinsulin was obtained by trifluoro-acetic acid, dimethyl sulfoxide and hydrochloric acid cleavage. The released proinsulin analog was confirmed by its molecular weight as determined by SDS-PAGE.

Keywords

Escherichia Coli Molecular Weight Dimethyl Fusion Protein Hydrochloric Acid 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Banting, F.G., and Best, C.H. (1922). J. Clin. Med. 7, 464–472.Google Scholar
  2. Bristow, A.F. (1993).Tibtech. 11, 301–305.Google Scholar
  3. Hellebust, H., Murby, M., Abrashmén, L., Uhlén, M., and Enfors, S.O. (1989) Bio/Technolog 7,165–168.Google Scholar
  4. Laemmli, U.K. (1970). Nature 227, 680–685.PubMedGoogle Scholar
  5. Martson, F.A.O. (1986). Bioch. J. 240, 1–12.Google Scholar
  6. Sambrook, J., Fritsch, E.F., and Maniatis, T.(1989), Molecular Cloning. A Laboratory Manual. Cold Spring Harbor Press.Google Scholar
  7. Sassenfeld, H.M., and Brewer, S.J. (1984). Bio/Thecnology 2, 76–81.Google Scholar
  8. Savige, W.E. and Fontana, A (1977). Meth.in Enzimol, 47, 459–469.Google Scholar
  9. Sayers J.R., Schmidt, W. and Eckstein, F. (1988). Nucl. Acids. Res. 16, 791–802.Google Scholar

Copyright information

© Chapman & Hall 1996

Authors and Affiliations

  • A. M. A. Nascimento
    • 1
    • 2
  • H. R. T. Souza
    • 3
  • M. S. Xavier
    • 3
  • J. E. Thiemann
    • 3
  • L. Vilela
    • 3
  • M. Mares-Guia
    • 2
    • 3
  1. 1.Departemento de Biologia GeralUFMGBelo HorizonteBrazil
  2. 2.Departamento de Bioquímica e ImunologiaUFMGBelo HorizonteBrazil
  3. 3.Biobras S.A, Distrito IndustrialMontes ClarosBrazil

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