Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Paper
  • Published:

Expression in Escherichia coli of Multiple Products from a Chimaeric Gene Fusion: Evidence for the Presence of Procaryotic Translational Control Regions within Eucaryotic Genes

Bio/Technology volume 3, pages 715–720 (1985)Cite this article

Abstract

Expression in Escherichia coli of a chimaeric gene fusion containing a porcine parvovirus (PPV) DNA fragment produced more than one size fusion protein. The introduction of a translation termination codon between the initiation codon and the PPV sequences of the gene fusion did not prevent the expression of the smaller fusion proteins. This suggests that the smaller products are the result of independent initiations of translation within PPV sequences rather than proteolysis of the full-length fusion protein. Three translation initiation sites were mapped to different segments of PPV DNA by protein sizing and deletion analysis. The sequences of these segments are presented.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Kleid, D.G., Yansura, D., Small, B., Dowbenko, D., Moore, D.M., Grubman, M.J., McKercher, P.O., Morgan, D.O., Robertson, B.H. and Bachrach, H.L. 1981. Cloned viral protein vaccine for foot and mouth disease:response in cattle and swine. Science 214: 1125–1129.

    Article  CAS  Google Scholar 

  2. Davis, A.R., Bos, T., Ueda, M., Nayak, D.P., Dowbenke, D. and Compans, R.W. 1983. Immune response to human influenza virus hemagglutinin expressed in Escherichia coli. Gene 21: 273–284.

    Article  CAS  Google Scholar 

  3. Watson, R.J., Weis, J.H., Salstrom, J.S. and Enquist, L.W. 1985. Expression of Herpes simplex virus type 1 and type 2 glycoprotein D genes using the Escherichia coli lac promoter, p. 327–352. In: Recombinant DNA Research and Viruses. Becker, Y. (ed), Martinus Nijhoff, The Hague.

    Chapter  Google Scholar 

  4. Casadaban, M. and Cohen, S.N. 1980. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J. Mol. Biol. 138: 179–207.

    Article  CAS  Google Scholar 

  5. Mengling, W.L. and Cultip, R.C. 1975. Pathogenesis of in utero infection: Experimental infection of five-week old porcine fetuses with porcine parvovirus. Am. J. Vet. Res. 36: 1173–1177.

    Google Scholar 

  6. Tattersall, P. 1978. Parvovirus protein structure and virion maturation, p. 53–72. In: Replication of Mammalian Parvoviruses. Tattersall, P. J. and Ward, D. C. (eds), Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

    Google Scholar 

  7. Berns, K.I. and Hauswirth, W.W. 1978. Parvovirus DNA structure and replication, p. 13–31. In: Replication of Mammalian Parvoviruses. Tattersall, P. J. and Ward, D. C. (eds.), Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

    Google Scholar 

  8. Moliter, T.W., Joo, H.S. and Collett, M.S. 1983. Porcine parvovirus: virus purification and structural and antigenic properties of virion polypeptides. J. Virol. 45: 842–854.

    Google Scholar 

  9. Astell, C.R., Thompson, M., Merchlinsky, M. and Ward, D.C. 1983. The complete DNA sequence of minute virus of mice, an autonomous parvovirus. Nucl. Acids Res. 11: 999–1018.

    Article  CAS  Google Scholar 

  10. Pintel, D., Dadachanji, D., Astell, C.R. and Ward, D.C. 1983. The genome of minute virus of mice, an autonomous parvovirus, encodes two overlapping transcription units. Nucl. Acids Res. 11: 1019–1038.

    Article  CAS  Google Scholar 

  11. Smith, S. and Halling, S.M. 1984. Expression of canine parvovirus-β-galactosidase fusion proteins in Escherichia coli. Gene 29: 263–269.

    Article  CAS  Google Scholar 

  12. Moliter, T.W., Joo, H.S. and Collett, M.S. 1984. Porcine parvovirus DNA: Characterization of the genomic and replicate form of two virus isolates. Virology 137: 241–254.

    Article  Google Scholar 

  13. Paradiso, P.R., Williams, K.R. and Costantino, R.L. 1984. Mapping of the amino terminus of the H-1 parvovirus major capsid protein. J. Virol. 52: 77–81.

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Watson, R.J., Weis, J.H., Salstrom, J.S. and Enquist 1982. Herpes Simplex virus type-1 glycoprotein D gene: Nucleotide sequence and expression in Escherichia coli. Science 218: 381–384.

    Article  CAS  Google Scholar 

  15. Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.

    Article  CAS  Google Scholar 

  16. Itakura, K., Hirose, T., Crea, R., Riggs, A.D., Heyneker, H.L., Bolivar, F. and Boyer, H.W. 1977. Expression in Escherichia coli of a chemically synthesized gene for the hormone somatostatin. Science 198: 1056–1063.

    Article  CAS  Google Scholar 

  17. Shine, J. and Dalgarno, L. 1975. Determinant of cistron specificity in bacterial ribosomes. Nature 254: 34–38.

    Article  CAS  Google Scholar 

  18. Steitz, J.A. 1979. Genetic signals and nucleotide sequences in messenger RNA, p. 349–399. In: Biological Regulation and Development.—1. Gene Expression. Goldberger, R. F. (ed). Plenum Press, New York.

    Chapter  Google Scholar 

  19. Chang, C.Y., Erlich, H.A., Gunsalus, R.P., Nunberg, J.H., Kaufman, R.J., Schimke, R.T. and Cohen, S.N. 1980. Initiation of protein synthesis in bacteria at a translational start codon of mammalian cDNA: Effects of the preceding nucletide sequence. Proc. Natl. Acad. Sci. USA 77: 1442–1446.

    Article  CAS  Google Scholar 

  20. Garapin, A.C., Colbere-Garpin, F., Cohen-Solal, M., Horodniceanu, F. and Kourilsky, P. 1981. Expression of herpes simplex virus type I thymidine kinase gene in Escherichia coli. Proc. Natl. Acad. Sci. USA 78: 815–819.

    Article  CAS  Google Scholar 

  21. Guarente, L., Lauer, G., Roberts, J.M. and Ptashne, M. 1980. Improved methods for maximizing expression of a cloned gene: a bacterium that synthesizes rabbit β-globin. Cell 20: 543–553.

    Article  CAS  Google Scholar 

  22. Cohen, S.N., Chang, A.C.Y. and Hsu, L. 1972. Non-chromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli. Proc. Natl. Acad. Sci. USA 69: 2110–2114.

    Article  CAS  Google Scholar 

  23. Miller, J.H. 1972. Experiments in Molecular Genetics, p. 431–433. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

    Google Scholar 

  24. Birnboim, H.C. and Doly, J. 1979. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucl. Acids Res. 7: 1513.

    Article  CAS  Google Scholar 

  25. Maniatis, T., Fritsch, E.F. and Sambrook, J. 1982. Molecular Cloning A Laboratory Manual, p. 93–94. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

    Google Scholar 

Download references

Author information

Author notes

  1. Susan Smith

    Present address: Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

Authors and Affiliations

  1. Molecular Genetics, Inc., 10320 Bren Road East, Minnetonka, MN

    Shirley M. Halling

Authors
  1. Shirley M. Halling
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Susan Smith
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Halling, S., Smith, S. Expression in Escherichia coli of Multiple Products from a Chimaeric Gene Fusion: Evidence for the Presence of Procaryotic Translational Control Regions within Eucaryotic Genes. Nat Biotechnol 3, 715–720 (1985). https://doi.org/10.1038/nbt0885-715

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt0885-715

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing