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Applied and Environmental Microbiology, August 1998, p. 2763-2769, Vol. 64, No. 8
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Use of the Lactococcal nisA Promoter To Regulate Gene Expression in Gram-Positive Bacteria: Comparison of Induction Level and Promoter Strength

Zehava Eichenbaum,1 Michael J. Federle,1 Diana Marra,1 Willem M. de Vos,2 Oscar P. Kuipers,2 Michiel Kleerebezem,2 and June R. Scott1,*

Department of Microbiology and Immunology, Emory University Health Sciences Center, Atlanta, Georgia 30322,1 and Department of Biophysical Chemistry, Section of Microbial Ingredients, Netherlands Institute for Dairy Research (NIZO), 6710 BA Ede, The Netherlands2

Received 13 March 1998/Accepted 14 May 1998

We characterized the regulated activity of the lactococcal nisA promoter in strains of the gram-positive species Streptococcus pyogenes, Streptococcus agalactiae, Streptococcus pneumoniae, Enterococcus faecalis, and Bacillus subtilis. nisA promoter activity was dependent on the proteins NisR and NisK, which constitute a two-component signal transduction system that responds to the extracellular inducer nisin. The nisin sensitivity and inducer concentration required for maximal induction varied among the strains. Significant induction of the nisA promoter (10- to 60-fold induction) was obtained in all of the species studied at a nisin concentration just below the concentration at which growth is inhibited. The efficiency of the nisA promoter was compared to the efficiencies of the Spac, xylA, and lacA promoters in B. subtilis and in S. pyogenes. Because nisA promoter-driven expression is regulated in many gram-positive bacteria, we expect it to be useful for genetic studies, especially studies with pathogenic streptococci in which no other regulated promoters have been described.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Emory University Health Sciences Center, Atlanta, GA 30322. Phone: (404) 727-0402. Fax: (404) 727-8999. E-mail: Scott{at}microbio.emory.edu.

Applied and Environmental Microbiology, August 1998, p. 2763-2769, Vol. 64, No. 8
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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