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, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Nakamura, K. and Inouye, M. 1982. Construction of versatile expression cloning vehicles using the lipoprotein gene of Escherichia coli. EMBO J. 1: 771–775.
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.
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.
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.
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.
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.
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.
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.
Anderson, C.W., Braum, P.R. and Gestland, R.F. 1973. Processing of adenovirus 2-induced proteins. J. Virol. 12: 241–252.
Author information
Authors and Affiliations
Rights 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
Issue Date:
DOI: https://doi.org/10.1038/nbt0184-81