Method for controlling the gene expression in lactic acid bacter

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...

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4352523, 4353201, 435462, 435463, 435465, 435471, 536 232, 536 241, C12P 2106, C12N 1500, C12N 120, C07H 2104

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059142480

ABSTRACT:
The present invention relates to a method for the controlled expression, in a lactic acid bacterium, of a DNA fragment containing one or more genes coding for a desired characteristic, wherein the DNA fragment is under the control of a promoter for a microbial gene which codes for an antimicrobial peptide, and the gene or genes are brought to expression on the DNA fragment by the addition of a suitable inducing factor for the transcription activation. The promoter is preferably the nisA promoter from Lactococcus lactis. The inducer is preferably acceptable for food products, and more preferentially is nisin or derivatives thereof. The invention also relates to a method for the production of proteins or RNA, as well as to a method for the preparation of dairy products using the expression system according to the invention. The invention finally relates to lactic acid bacteria and expression vectors for use in the method according to the invention.

REFERENCES:
Lakshmidevi et al., Molecular Characterization of Promoters of the Lactococcus lactis subsp. cremoris Temperate Bacteriophage BK5-T and Identification of a Phage Gene Implicated in the Regulation of Promoter Activity, Applied and Environmental Microbiology, vol. 56, No. 4, Apr. 1990, pp. 934-942, contents page.
Kirsch et al., The Use of B-Galactosidase Gene Fusions to Screen for Antibacterial Antibiotics, An International Journal Devoted to Research on Bioactive Microbial Products, The Journal of Antibiotics, vol. 44, No. 2, Feb. 1991, pp. 210-217, contents page.
van der Meer et al., Characterization of the Lactococcus lactis Nisin A Operon Genes nisP, Encoding a Subtilisin-Like Serine Protease Involved in Precursor Processing, and nisR, Encoding a Regulatory Protein Involved in Nisin Biosynthesis, Journal of Bacteriology, vol. 175, No. 9, May 1993, pp. 2578-2588.
Shearman et al., Autolytic Lactococcus lactis Expressing a Lactococcal Bacteriophage Lysin Gene, Bio/Technology, vol. 10, No. 2, Feb. 1992, pp. 196-199, contents page.
Kuroda et al., Molecular Cloning and Sequencing of the Upstream Region of the Major Bacillus subtilis Autolysin Gene: a Modifier Protein Exhibiting Sequence Homology to the Major Autolysin and the spoIID Product, Journal of General Microbiology, vol. 138, Part 6, pp. 1067-1076.
Kuipers et al., Characterization of the Nisin Gene Cluster nisABTCIPR of Lactococcus lactis Requirement of Expression of the nisA and nisI Genes for Development of Immunity, European Journal of Biochemistry, vol. . 216, No. 1, Aug. 1993, pp. 281-291.
Kuipers, Oscar P. et al., "Autoregulation of Nisin Biosynthesis in Lactococcus lactis by Signal Transduction," J Biol Chem, Nov. 10, 1995, vol. 270, No. 45, pp. 27299-27304.
Engelke, G. et al., "Regulation of Nisin Biosynthesis and Immunity in Lactococcus lactis 6F3" Appl Environ Microbiol, Mar. 1994, vol. 60, No. 3, pp. 814-825.

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