The Expression of Human Papillomavirus Type 18 E6 Protein in Bacteria and the Production of Anti-E6 Antibodies Free

Abstract

Summary

Human papillomavirus type 18 (HPV-18) has recently been closely linked with human malignant cervical lesions. The early region of the genome of the related bovine papillomavirus (BPV) has been shown to be important for the production of the transformed phenotype. BPV E6 has been shown to be a transforming protein. We report the primary structure of the HPV-18 E6 open reading frame and its predicted amino acid sequence. Both E6 protein and E6-β-galactosidase fusion protein were synthesized in bacteria. Antisera were raised against the E6-β-galactosidase fusion protein and against an E6 N-terminal peptide which was 14 amino acids long. We show that these antisera reacted on Western blots against E6 synthesized in bacteria. The HPV E6 antigen and antibodies described here will be useful in understanding HPV expression and its association with human malignancies and may also be diagnostically useful.

Keyword(s): antibodies , DNA sequence , expression and HPV
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1986-09-01
2024-03-29
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References

  1. Androphy E. J., Schiller J. T., Lowy D. R. 1985; Identification of the protein encoded by the E6 transforming gene of bovine papillomavirus. Science 230:442–445
    [Google Scholar]
  2. Boshart M., Gissmann L., Ikenberg H., Kleinheinz A., Scheurlen W., Zur Hausen H. 1984; A new type of papillomavirus DNA, its presence in genital cancer biopsies and in cell lines derived from cervical cancer. EMBO Journal 3:1151–1157
    [Google Scholar]
  3. Chang N. T., Huang J., Ghrayeb J., Mckinney S., Chanda P. K., Chang T. W., Putney S., Sarnagadharan M.G., Wong-Staal F., Gallo R. C. 1985; An HTLV-III peptide produced by recombinant DNA is immunoreactive with sera from patients with AIDS. Nature, London 315:151–154
    [Google Scholar]
  4. Crum C. P., Ikenberg H., Richart R. M., Gissmann L. 1984; Human papillomavirus type 16 and early cervical neoplasia. New England Journal of Medicine 310:880–883
    [Google Scholar]
  5. Gissmann L., Wolnick L., Ikenberg H., Koldovsky U., Schnurch H. G., Zur Hausen H. 1983; Human papillomavirus type 6 and 11 DNA sequences in genital and laryngeal papillomas and some cervical cancers. Proceedings of the National Academy of Sciences, U.S.A 80:560–563
    [Google Scholar]
  6. Ikenberg H., Gissmann L., Gross G., Grussendorf-Conen E., Zur Hausen H. 1983; Human papillomavirus type 16 related DNA in genital Bowen’s disease and Bowenoid papulosis. International Journal of Cancer 32:563–565
    [Google Scholar]
  7. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  8. Matlashewski G., Banks L., Pim D., Crawford L. 1986; Analysis of human p53 proteins and mRNA levels in normal and transformed cells. European Journal of Biochemistry 154:665–672
    [Google Scholar]
  9. Nelson L. M., Rothbard J. B., Shiurba A. R., Linke K. H., Schoolnik G. K., Clayton A. D. 1984; All eight unassigned reading frames of mouse mitochondrial DNA are expressed. EMBO Journal 3:3165–3175
    [Google Scholar]
  10. Rosenberg M., Ho Y., Shatzman A. 1983; The use of pKC30 and its derivatives for controlled expression of genes. Methods in Enzymology 101:123–138
    [Google Scholar]
  11. Ruther U., Muller-Hill B. 1983; Easy identification of cDNA clones. EMBO Journal 2:1791–1794
    [Google Scholar]
  12. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences, U.S.A 74:5463–5467
    [Google Scholar]
  13. Schiller J. T., Vass W. C., Lowy D. R. 1984; Identification of a second transforming region in bovine papillomavirus DNA. Proceedings of the National Academy of Sciences, U.S.A 81:7880–7884
    [Google Scholar]
  14. Schwarz E., Freese U. K., Gissmann L., Mayer W., Roggenbuck B., Stremlau A., Zur Hausen H. 1985; Structure and transcription of human papillomavirus sequences in cervical carcinoma cells. Nature, London 314:111–114
    [Google Scholar]
  15. Seedorf K., Krammer G., Durst M., Suhai S., Rowekamp M. 1985; Human papillomavirus type 16 DNA sequence. Virology 145:181–185
    [Google Scholar]
  16. Yang Y. C., Okayama H., Howley P. M. 1985; Bovine papillomavirus contains multiple transforming genes. Proceedings of the National Academy of Sciences, U.S.A 82:1020–1023
    [Google Scholar]
  17. Yee C., Krishnan-Hewlett I., Baker C. C., Schlegel R., Howley P. M. 1985; Presence and expression of human papillomavirus sequences in human cervical cell lines. American Journal of Pathology 119:361–366
    [Google Scholar]
  18. Zur Hausen H. 1985; Genital papillomavirus infections. In Viruses and Cancer pp. 83–90 Edited by Rigby P. W. J., Wilkie N. M. Cambridge: Cambridge University Press;
    [Google Scholar]
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