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Construction of a gene of the human tumor-associated antigen VNTR(MUC1) bound to streptavidin, its expression inEscherichia coli, and the study of properties of the hybrid protein

Abstract

A gene of human tumor-associated antigen VNTR(MUC1) bound to streptavidin, an expression plasmid, and a highly effective hybrid protein-producing strain were constructed. It was shown that the streptavidin leader peptide ensures effective secretion of the hybrid protein into the periplasmic space ofEscherichia coli cells. The hybrid protein was isolated in a homogeneous state and its immunogenic properties were studied.

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Abbreviations

KLH:

snail hemocyanine

SAV:

streptavidin

SAV(TR)22 :

the hybrid protein

TR:

a tandem repeat monomer

TyR:

bovine tyroglobulin

VNTR:

the MUC1 protein locus with a variable number of tandem repeats; prefix “d” in deoxyribooligonucleotides is omitted

References

  1. Finn, O.J., Jerome, K.R., Henderson, R.A., Pecher, G., Domenech, N., Magarian-Blander, J., and Barratt-Boyes, S.M.,Immunol. Rev., 1995, vol. 145, pp. 61–88.

    PubMed  Article  CAS  Google Scholar 

  2. Apostolopoulos, V., Lofthouse, S.A., Popovski, V., Chelvanayagam, G., Sandrin, M.S., and McKenzie, I.H.G.,Nature Biotechnol., 1998, vol. 15, pp. 276–280.

    Article  Google Scholar 

  3. Gendler, S.J., Lancaster, C.A., Taylor-Papadimitriou, J., Duhig, T., Peat, N., Burchell, J., Pemberton, L., Lalani, E.-N., and Wilson, D.,J. Biol. Chem., 1990, vol. 265, pp. 15286–15293.

    PubMed  CAS  Google Scholar 

  4. Apostolopoulos, V., and McKenzie, I.F.,Crit. Rev. Immunol., 1994, vol. 14, pp. 293–309.

    PubMed  CAS  Google Scholar 

  5. Lancaster, C.A., Peat, N., Duhig, T., Wilson, D., Taylor-Papadimitriou, J., and Gendler, S.J.,Biochem. Biophys. Res. Commun., 1990, vol. 173, pp. 1019–1029.

    PubMed  Article  CAS  Google Scholar 

  6. Pratt, W.S., Islam, I., and Swallow, D.M.,Ann. Hum. Genet., 1996, vol. 60, pp. 21–28.

    PubMed  Article  CAS  Google Scholar 

  7. Magarian-Blander, J., Ciborowski, P., Hsia, S., Watkins, S.C., and Finn, O.J.,J. Immunol., 1998, vol. 160, pp. 3111–3120.

    PubMed  CAS  Google Scholar 

  8. Ligtenberg, M.J., Vos, H.L., Gennissen, A.M., and Hilkens, J.,J. Biol. Chem., 1990, vol. 265, pp. 5573–5578.

    PubMed  CAS  Google Scholar 

  9. Griffiths, A.B., Burchell, J., Genoler, S., Lewis, A., Blight, K., Tilly, R., and Taylor-Papadimitriou, J.,Int. J. Cancer, 1987, vol. 40, pp. 319–327.

    PubMed  Article  CAS  Google Scholar 

  10. Burchell, J., Taylor-Papadimitriou, J., Boshell, M., Gendler, S., and Duhig, T.,Int. J. Cancer, 1989, vol. 44, pp. 691–696.

    PubMed  Article  CAS  Google Scholar 

  11. Barnd, D.L., Lan, M.S., Metzgar, R.S., and Finn, O.J.,Proc. Natl. Acad. Sci. USA, 1989, vol. 86, pp. 7159–7163.

    PubMed  Article  CAS  Google Scholar 

  12. Agarwal, B., Reddish, M.A., and Longenecker, B.M.,J. Immunol., 1996, vol. 157, pp. 2089–2095.

    Google Scholar 

  13. Jerom, K.R., Domenech, N., and Finn, O.J.,J. Immunol., 1993, vol. 151, pp. 1654–1662.

    Google Scholar 

  14. Apostolopoulos, V., Chelvanayagam, G., Xing, P.-X., and McKenzie, I.F.,J. Immunol., 1998, vol. 161, pp. 767–775.

    PubMed  CAS  Google Scholar 

  15. Apostolopoulos, V., Pietersz, G.A., Lovelend, B.E., Sandrin, M.S., and McKenzie, I.F.,Proc. Natl. Acad. Sci. USA, 1995, vol. 92, pp. 10128–10132.

    PubMed  Article  CAS  Google Scholar 

  16. Banchereau, J. and Steinman, R.M.,Nature, 1998, vol. 392, pp. 245–252.

    PubMed  Article  CAS  Google Scholar 

  17. Pecher, G. and Finn, O.J.,Proc. Natl. Acad. Sci. USA, 1996, vol. 93, pp. 1699–1704.

    PubMed  Article  CAS  Google Scholar 

  18. Gyllensten, U.,PCR Technology: Principles and Applications for DNA Amplification, Erlich, N.A., Ed., New York: Stockton, 1989, pp. 45–60.

    Google Scholar 

  19. Prince, J.T., McGrath, K.P., DiGirolamo, C.M., and Kaplan, D.L.,Biochemistry, 1995, vol. 34, pp. 10879–10885.

    PubMed  Article  CAS  Google Scholar 

  20. Kam, J.L., Regimbald, L.H., Hilgers, J.H.M., Hoffman, P., Krantz, M.J., Longenecker, M., and Hugh, J.C.,Cancer Res., 1998, vol. 58, pp. 5577–5581.

    PubMed  CAS  Google Scholar 

  21. Argarana, C.E., Kuntz, I.D., Birken, S., Alex, R., and Cantor, C.R.,Nucleic Acids Res., 1986, vol. 14, pp. 1871–1882.

    PubMed  Article  CAS  Google Scholar 

  22. Weber, P.C., Pantoliano, M.W., and Thompson, L.D.,Biochemistry, 1992, vol. 31, pp. 9250–9254.

    Google Scholar 

  23. Olejnik, J., Sonar, S., Krzymanska-Olejnik, E., and Rothschild, K.,Proc. Natl. Acad. Sci. USA, 1995, vol. 92, pp. 7590–7594.

    PubMed  Article  CAS  Google Scholar 

  24. Sano, T., Vajda, S., and Cantor, C.,J. Chromatogr. B, 1998, vol. 715, pp. 85–91.

    Article  CAS  Google Scholar 

  25. Gul’ko, L.B., D’yakov, N.A., Okorokova, N.A., Veiko, V.P., and Debabov, V.G.,Biotechnologiya, 1999, no. 4, pp. 3–8.

  26. Dyakov, N.A., Gul’ko, L.B., Okorokova, N.A., and Veiko, V.P.,9th European Congr. on Biotechnology, 1999, ECB9/2804.

  27. Veiko, V.P., Gul’ko, L.B., Okorokova, N.A., D’yakov, N.A., and Debabov, V.G.,Bioorg. Khim., 1999, vol. 25, pp. 185–189.

    Google Scholar 

  28. Wei, R.D.,Methods Enzymol., 1970, vol. 18A, pp. 424–427.

    Article  Google Scholar 

  29. Lloyd, K.D., Burchell, J., Kudryashov, V., Yin, B.W., and Taylor-Papadimitriou, J.,J. Biol. Chem., 1996, vol. 271, pp. 33325–33334.

    PubMed  Article  CAS  Google Scholar 

  30. Sambrook, J., Fritsch, E.F., and Maniatis, T.,Molecular Cloning: A Laboratory Manual., 2nd ed., Cold Spring Harbor, NY: Cold Spring Harbor Lab., 1989, pp. 1.22, 1.53, 1.74, 17.32.

    Google Scholar 

  31. Veiko, V.P., Ratmanova, K.I., Osipov, A.S., Bulenkov, M.T., and Pugachev, V.V.,Bioorg. Khim., 1991, vol. 17, pp. 625–629.

    Google Scholar 

  32. Sanger, F., Nicklen, S., and Coulson, A.R.,J. Mol. Biol., 1975, vol. 94, pp. 441–446.

    PubMed  Article  CAS  Google Scholar 

  33. Veiko, V.P., Osipov, A.S., Shekhter, I.I., Bulenkov, M.T., Ratmanova, K.I., Gul’ko, L.B., Chibiskova, N.A., Errais, L.L., Derevshchikova, E.B., and Debabov, V.G.,Bioorg. Khim., 1995, vol. 21, pp. 354–358.

    PubMed  CAS  Google Scholar 

  34. Laemmli, U.,Nature, 1970, vol. 227, pp. 680–685.

    PubMed  Article  CAS  Google Scholar 

  35. Hofmann, K., Wood, S.W., Brinton, C.C., Montibeller, J.A., and Finn, M.,Proc. Natl. Acad. Sci. USA, 1980, vol. 77, pp. 4666–4668.

    PubMed  Article  CAS  Google Scholar 

  36. Bradford, M.M.,Anal. Biochem., 1976, vol. 72, p. 248.

    PubMed  Article  CAS  Google Scholar 

  37. Tijssen, P.,Laboratory Techniques in Biochemistry and Molecular Biology, Burdon, R.H. and van Knippenberg, P.H., Eds., Amsterdam: Elsevier, 1986, vol. 15, pp. 258–270.

    Google Scholar 

  38. Ionov, Yu. V. and Yurin, V.L.,Immunologiya, 1992, vol. 4, pp. 57–60.

    Google Scholar 

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Correspondence to V. P. Veiko.

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Gul’ko, L.B., Pavlova, O.V., D’yakov, N.A. et al. Construction of a gene of the human tumor-associated antigen VNTR(MUC1) bound to streptavidin, its expression inEscherichia coli, and the study of properties of the hybrid protein. Russ J Bioorg Chem 26, 381–389 (2000). https://doi.org/10.1007/BF02758665

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  • DOI: https://doi.org/10.1007/BF02758665

Key words

  • Escherichia coli
  • hybrid protein
  • MUC1
  • streptavidin
  • tumor-associated antigen
  • VNTR