Abstract
A novel expression vector based on the rmf promoter (Wada et al. Proc. Natl. Acad. Sci. USA (1995) 87: 2657–2661) was developed. Using lacZ gene as a model, protein production depending on cell growth phase and growth rate was found. In addition, by subjecting the recombinant cells to temperature downshift and substrate famine, the strain was capable of producing 35 000–40 000 units of β-galactosidase. These characteristics indicate the potential application of this expression vector to bioprocessing such as protein production in continuous or batch fermentation, biotransformation using entrapped cells, and in situ bioremediation.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Cashel M, Gentry DR, Hernandez VJ, Ninella D (1996) The stringent response. In: Neiehardt FC, Curtiss R, Ingraham JL, Lin ECC, Low KB, Magasanik B, Rezikoff WS, Riley M, Schaechter M, Umbarger HE, eds. Escherichia coli and Salmonella: Cellular and Molecular Biology. Washington, DC: American Society for Microbiology, pp. 1458–1496.
Chao YP, Juang TY, Chern JT, Lee CK (1999) Production of D-p-hydroxyphenylglycine by N-carbamoyl-D-amino acid amidohydrolase-overproducing Escherichia coli strains. Biotechnol. Prog. 15: 603–607.
Churchward G, Belin D, Nagamine Y (1984) A pSC101-derived plasmid which shows no sequence homology to other commonly used cloning vectors. Gene 31: 165–171.
Furste JP, Pansegrau W, Frank R, Blocker H, Scholz P, Bagdasarian M, Lanka E (1986) Molecular cloning of the plasmid RP4 primase region in a multi-host-range tacP expression vector. Gene 48: 119–131.
Han SJ, Chang HN, Lee J (1998) Fed-batch cultivation of an oxygen-dependent inducible promoter system, the nar promoter in Escherichia coli with an inactivated nar operon. Biotechnol. Bioeng. 59: 400–406.
Ishihama A (1997) Adaptation of gene expression in stationary phase bacteria. Curr. Opin. Gene. Develop. 7: 582–588.
Makrides SC (1996) Strategies for achieving high-level expression of genes in Escherichia coli. Microbiol. Rev. 60: 512–538.
Matin A (1994) Starvation promoters of Escherichia coli: their function, regulation, and use in bioprocessing and bioremediation. Ann. N.Y. Acad. Sci. 721: 277–291.
Miksch G, Fiedler E, Dobrowolski P., Friehs K (1997) The kil gene of the ColE1 plasmid of Escherichia coli controlled by a growth-phase-dependent promoter mediates the secretion of a heterologous periplasmic protein during the stationary phase. Arch. Microbol. 167: 143–150.
Miller JH (1972) Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
Neidhardt FC, Ingraham JL, Schaechte M (1990) Physiology of the Bacterial Cell: A Molecular Approach. Sunderland, MS: Sinauer Associates, Inc.
Parales RE, Harwood CS (1993) Construction and use of a new broad-host-range lacZ transcriptional fusion vector, pHRP309, for gram negative bacteria. Gene 133: 23–30.
Tanaka K, Takayanagi Y, Fujita N, Ishihama A, Takahashi H (1993) Heterogeneity of the principle σ factor in Escherichia coli: the rpoS gene product, σ38, is a second principle factor of RNA polymerase in stationary-phase Escherichia coli. Proc. Natl. Acad. Sci. USA. 90: 3511–3515.
Thomas JG, Ayling A, Baneyx F (1997) Molecular chaperones, folding catalysts, and the recovery of active recombinant proteins from E. coli. Appl. Biochem. Biotechnol. 66: 197–238.
Tunner JR, Robertson CR, Schippa S, Matin A (1992) Use of glucose starvation to limit growth and induce protein production in Escherichia coli. Biotechnol. Bioeng. 40: 271–279.
Vasina JA, Baneyx F (1996) Recombinant protein expression at low temperatures under the transcriptional control of the major Escherichia coli cold shock promoter cspA. Appl. Environ. Microbiol. 62: 1444–1447.
Wada A, Igarashi K, Yoshimura S, Aimoto S, Ishihama A (1995) Ribosome modulation factor: stationary growth phase-specific inhibitor of ribosome functions from Escherichia coli. Biochem. Biophys. Res. Comm. 214: 410–417.
Wada A, Yamazaki Y, Fujita N, Ishihama A (1990) Structure and probable genetic location of a ‘ribosome modulation factor’ associated with 100S ribosomes in stationary-phase Escherichia coli cells. Proc. Natl. Acad. Sci. USA 87: 2657–2661.
White D (1995) The Physiology and Biochemistry of Prokaryotes. New York: Oxford University Press.
Yamagishi M, Matsushima H, Wada A, Sakagami M, Fujita N, Ishihama A (1993) Regulation of the Escherichia coli rmf gene encoding the ribosome modulation factor: growth phase-and growth rate-dependent control. EMBO J. 12: 625–630.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Chao, YP., Wen, CS., Chiang, CJ. et al. Construction of the expression vector based on the growth phase- and growth rate-dependent rmf promoter: use of cell growth rate to control the expression of cloned genes in Escherichia coli. Biotechnology Letters 23, 5–11 (2001). https://doi.org/10.1023/A:1026775906596
Issue Date:
DOI: https://doi.org/10.1023/A:1026775906596