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J Bacteriol, February 1998, p. 571-577, Vol. 180, No. 3
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Genetic Analysis, Using P22 Challenge Phage, of the Nitrogen Activator Protein DNA-Binding Site in the Klebsiella aerogenes put Operon

Li-Mei Chen,^1, Thomas J. Goss,² Robert A. Bender,² Simon Swift,³ and Stanley Maloy^1,*

Department of Biology, University of Michigan, Ann Arbor, Michigan 48109-1048²; Department of Applied Biochemistry and Food Science, University of Nottingham, Sutton Bonington Campus, Leicestershire LE12 5RD, United Kingdom³; and Department of Microbiology, University of Illinois, Urbana, Illinois 61801¹

Received 11 July 1997/Accepted 24 November 1997

The nac gene product is a LysR regulatory protein required for nitrogen regulation of several operons from Klebsiella aerogenes and Escherichia coli. We used P22 challenge phage carrying the put control region from K. aerogenes to identify the nucleotide residues important for nitrogen assimilation control protein (NAC) binding in vivo. Mutations in an asymmetric 30-bp region prevented DNA binding by NAC. Gel retardation experiments confirmed that NAC specifically binds to this sequence in vitro, but NAC does not bind to the corresponding region from the put operon of Salmonella typhimurium, which is not regulated by NAC.

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^* Corresponding author. Mailing address: Department of Microbiology, University of Illinois, B103 Chemical and Life Sciences Laboratory, 601 S. Goodwin Ave., Urbana, IL 61801. Phone: (217) 333-3122. Fax: (217) 244-6697. E-mail: s-maloy{at}uiuc.edu.

Present address: Department of Microbiology, SUNY, Stony Brook, N.Y.

This article has been cited by other articles:

• Muse, W. B., Bender, R. A. (1998). The nac (Nitrogen Assimilation Control) Gene from Escherichia coli. J. Bacteriol. 180: 1166-1173 [Abstract] [Full Text]  
• Pomposiello, P. J., Janes, B. K., Bender, R. A. (1998). Two Roles for the DNA Recognition Site of the Klebsiella aerogenes Nitrogen Assimilation Control Protein. J. Bacteriol. 180: 578-585 [Abstract] [Full Text]