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:

A Novel Prophage Independent trp Regulated Lambda PL Expression System

Bio/Technology volume 4, pages 802–808 (1986)Cite this article

Abstract

A new lambda PL/cItrp regulation system was constructed which places the lambda cI repressor gene under the control of the tryptophan promoter/operator. This system allows prophage and temperature independent gene expression of lambda promoter plasmids. Protein synthesis is induced by blocking the production of lambda cI represser molecules through the addition to the growth medium of inexpensive supplements rich in tryptophan. Two cItrp constructs were placed on F′–factors for easy transfer into any suitable E. coli strain. The system should be useful for large scale production in E. coli of enzymes or other low added–value proteins.

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. Brosius, J. 1984. Toxicity of an overproduced foreign gene product in Escherichia coli and its use in plasmid vectors for the selection of transcription terminators. Gene 27:161–172.

    Article  CAS  Google Scholar 

  2. Carrier, M.J., Nugent, M.E., Tacon, W.C.A., and Primrose, S.B. 1983. High expression of cloned genes in E. coli and its consequences. Trends Biotechnol. 1:109–113.

    Article  CAS  Google Scholar 

  3. Remaut, E., Tsao, H., and Fiers, W. 1983. Improved plasmid vectors with a thermoinducible expression and temperature-regulated run-away replication. Gene 22:103–113.

    Article  CAS  Google Scholar 

  4. Shimatake, H. and Rosenberg, M. 1982. Purified λ regulatory protein cII positively activates promoters for lysogenic development. Nature 292:128–132.

    Article  Google Scholar 

  5. Mieschendahl, M. and Müller-Hill, B. 1985. F′-coded, temperature-sensitive λ cI857 represser gene for easy construction and regulation of λ promoter-dependent expression systems. J. Bacteriol. 164:1366–1369.

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Sanger, F., Coulson, A.R., Hong, G.F., Hill, D.F., and Petersen, G.B. 1982. Nucleotide sequence of bacteriophage λ DNA. J. Mol. Biol. 162:729–773.

    Article  CAS  Google Scholar 

  7. Koenen, M., Rüther, U., and Müller-Hill, B. 1982. Immunoenzymatic detection of expressed gene fragments cloned in the lacZ gene of E. coli. EMBO J. 1:509–512.

    Article  CAS  Google Scholar 

  8. Langley, K.E., Villarejo, M.R., Fowler, A.V., Zamenhof, P.J., and Zabin, I. 1975. Molecular basis of β-galactosidase alpha-complementation. Proc. Natl. Acad. Sci. USA 72:1254–1257.

    Article  CAS  Google Scholar 

  9. Mieschendahl, M. 1981. Isolation and analysis of λ N−-lacZ+, λ rex−-lacZ+, and lacY+-lacZ+ gene fusions in E. coli. Thesis, Universität zu Köln

    Google Scholar 

  10. Russel, D.R. and Bennett, G.N. 1982. Cloning of small DNA fragments containing the Escherichia coli tryptophan operon promoter and operator. Gene 17:9–18.

    Article  CAS  Google Scholar 

  11. Hull, E.P., Spencer, M.E., Wood, D., and Guest, J.R. 1983. Nucleotide sequence of the promoter region of the citrate synthase gene (gltA) of Escherichia coli. FEBS Lett. 156:366–370.

    Article  CAS  Google Scholar 

  12. Mieschendahl, M., Grießer, H.-W., and Müller-Hill, B. 1981. λ immunity phase shift in a λ N−-lacZ+ fusion. Mol. Gen. Genet. 183:202–204.

    Article  CAS  Google Scholar 

  13. Miller, J.H. 1972. Experiments in Molecular Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.

    Google Scholar 

  14. Guest, J.R. 1981. Hybrid plasmid containing the citrate synthase gene (gltA) of Escherichia coli K12. J. Gen. Microbiol. 124:17–23.

    CAS  PubMed  Google Scholar 

  15. Masui, Y., Mizumo, T., and Inouye, M. 1984. Novel high-level expression cloning vehicles: 104-fold amplification of Escherichia coli minor protein. Bio/Technology 2:81–85.

    CAS  Google Scholar 

  16. Lugtenberg, B. and van Alphen, L. 1983. Molecular architecture and functioning of the outer membrane of Escherichia coli and other gram-negative bacteria. Biochim. Biophys. Acta 737:51–115.

    Article  CAS  Google Scholar 

  17. Gabain, A.V. and Bujard, H. 1979. Interaction of Escherichia coli RNA polymerase with promoters of several coliphages and plasmid DNA. Proc. Natl. Acad. Sci. USA 76:189–193.

    Article  Google Scholar 

  18. Zabin, I. and Fowler, A.V. 1978. β-Galactosidase, the lactose permease protein, and thiogalactoside transacetylase, p. 89–121. In: The Operon. J. H. Miller and W. S. Reznikoff (eds.). Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.

    Google Scholar 

  19. Tacon, W., Carey, N., and Emtage, S. 1980. The construction and characterisation of plasmid vectors suitable for the expression of all DNA phases under the control of the E. coli tryptophan promoter. Mol. Gen. Genet. 177:427–438.

    Article  CAS  Google Scholar 

  20. Itakura, K., Hirose, T., Crea, 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 

  21. De Boer, H.A., Comstock, L.J., and Vasser, M. 1983. The tac promoter: a functional hybrid derived from the trp and lac promoters. Proc. Natl. Acad. Sci. USA 80:21–25.

    Article  CAS  Google Scholar 

  22. Boidel, W., Simonis, M., Töpert, M., and Siewert, G. 1982. Recombinant plasmids with genes for the biosynthesis of alkaline phosphatase of Escherichia coli. Molec. Gen. Genet. 185:510–512.

    Article  Google Scholar 

  23. Waldman, A.S., Haeusslein, E., and Milman, G. 1983. Purification and characterization of herpes simplex (type 1) thymidine kinase produced in Escherichia coli by a high efficiency expression plasmid utilizing a λ PL promoter and a cI857 temperature-sensitive repressor. J. Biol. Chem. 258:11571–11575.

    CAS  PubMed  Google Scholar 

  24. Zabeau, M. and Stanley, K.K. 1982. Enhanced expression of cro-β-galactosidase fusion proteins under the control of the PR promoter of bacteriophage λ. EMBO J. 1:1217–1224.

    Article  CAS  Google Scholar 

  25. Reichardt, L.F. 1975. Control of bacteriophage lambda repressor synthesis: regulation of the maintenance pathway by the cro and cI product. J. Mol. Biol. 93:289–309.

    Article  CAS  Google Scholar 

  26. Rüther, U., Koenen, M., Otto, K., and Müller-Hill, B. 1981. pUR222, a vector for cloning and rapid chemical sequencing of DNA. Nucl. Acids Res. 9:4087–4098.

    Article  Google Scholar 

  27. Maniatis, T., Fritsch, E.F., and Sambrook, J. 1982. Molecular Cloning, A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.

    Google Scholar 

  28. Rosenberg, M., Ho, Y.-S., and Shatzman, A. 1983. The use of pKC30 and its derivatives for controlled expression of genes. Methods Enzymol. 101:123–138.

    Article  CAS  Google Scholar 

Download references

Author information

Author notes

  1. Thomas Petri

    Present address: Schering AG, D-1000, Berlin, 65, FRG

  2. Urs Hänggi

    Present address: Henkel KG, D-4000, Düsseldorf, 1, FRG

Authors and Affiliations

  1. Battelle-Institut, Am Römerhof 35, D-6000, Frankfurt/M, 90, Federal Republic of Germany

    Martin Mieschendahl

Authors
  1. Martin Mieschendahl
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Thomas Petri
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Urs Hänggi
    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

Mieschendahl, M., Petri, T. & Hänggi, U. A Novel Prophage Independent trp Regulated Lambda PL Expression System. Nat Biotechnol 4, 802–808 (1986). https://doi.org/10.1038/nbt0986-802

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt0986-802

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