Skip to main content
Log in

Novel High-level Expression Cloning Vehicles: 104-fold Amplification of Escherichia coli Minor Protein

  • Research Paper
  • Published:
Bio/Technology Submit manuscript

Abstract

Versatile, high-level expression cloning vehicles, pINIII, have been constructed. A foreign DNA fragment can be inserted in any one of the three reading frames at the unique EcoRI, HindIII, or BamHI site immediately after the initiation codon. The cloned foreign gene is under the control of both the lpp promoter and the lac promoter-operator. The expression of the gene is regulated by the lac repressor produced by the same vehicles. Using a pINIII vehicle EnvZ protein, a very minor regulatory protein of Escherichia coli was amplified more than 104-fold, becoming 30% of total cellular protein.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Nakamura, K. and Inouye, M. 1982. Construction of versatile expression cloning vehicles using the lipoprotein gene of Escherichia coli. EMBO J. 1: 771–775.

    Article  CAS  Google Scholar 

  2. Reznikoff, W.S. and Abelson, J., 1978. The lac Promoter. p. 221–243. In: The Operon. J.H. Miller and W.S. Reznikoff (eds.), Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.

    Google Scholar 

  3. Mizuno, T., Chou, M.-Y. and Inouye, M. 1983. A comparative study on the genes for three porins of the Escherichia coli outer membrane: DNA sequence of osmoregulated ompC gene. J. Biol. Chem. 258: 6932–6940.

    CAS  PubMed  Google Scholar 

  4. Movva, N.R., Nakamura, K. and Inouye, M. 1980. Regulatory region of the gene for the OmpA protein, a major membrane protein of Escherichia coli. J. Biol. Chem. 143: 317–328.

    CAS  Google Scholar 

  5. Nakamura, K., Masui, Y. and Inouye, M. 1982. Use of a lac promoter-operator fragment as a transcriptional control switch for expression of the constitutive lpp gene in Escherichia coli. J. Mol. Appl. Genet. 1: 289–299.

    CAS  PubMed  Google Scholar 

  6. Mizuno, T., Wurtzel, E.T. and Inouye, M. 1982. Osmoregulation of gene expression. II. DNA sequences of the envZ gene of the ompB operon of Escherichia coli and characterization of its gene product. J. Biol. Chem. 257: 13692–13698.

    CAS  PubMed  Google Scholar 

  7. Masui, Y., Coleman, J. and Inouye, M. 1983. Multipurpose expression cloning vehicles in Escherichia coli. In: Experimental Manipulation of Gene Expression. M. Inouye (ed.), Academic Press, New York. In press.

    Google Scholar 

  8. Wurtzel, E.T., Movva, N.R., Ross, F. and Inouye, M. 1981. Two-step cloning of the Escherichia coli regulatory gene ompB, employing phage Mu. J. Mol. Appl. Gent. 1: 61–69.

    CAS  Google Scholar 

  9. Anderson, C.W., Braum, P.R. and Gestland, R.F. 1973. Processing of adenovirus 2-induced proteins. J. Virol. 12: 241–252.

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Masui, Y., Mizuno, T. & Inouye, M. Novel High-level Expression Cloning Vehicles: 104-fold Amplification of Escherichia coli Minor Protein. Nat Biotechnol 2, 81–85 (1984). https://doi.org/10.1038/nbt0184-81

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt0184-81

  • Springer Nature America, Inc.

This article is cited by

Navigation