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Journal of Bacteriology, May 2001, p. 3169-3175, Vol. 183, No. 10
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.10.3169-3175.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Promoter Cloning in the Radioresistant Bacterium Deinococcus radiodurans

Rob Meima,1,dagger Heather M. Rothfuss,1 Lindy Gewin,2 and Mary E. Lidstrom1,3,*

Department of Chemical Engineering,1 Cellular and Molecular Biology Program,2 and Department of Microbiology,3 University of Washington, Seattle, Washington 98195

Received 22 June 2000/Accepted 26 February 2001

Deinococcus radiodurans is a highly radiation-resistant bacterium that is classed in a major subbranch of the bacterial domain. Since very little is known about gene expression in this bacterium, an initial study of promoters was undertaken. In order to isolate promoters and study promoter function, a series of integrative vectors for stable chromosomal insertion in D. radiodurans were developed. These vectors are based on Escherichia coli replicons that are unable to replicate autonomously in D. radiodurans and carry homologous sequences for replacement recombination in the D. radiodurans chromosome. The resulting integration vectors were used to study expression of reporter genes fused to a number of putative promoters that were amplified from the D. radiodurans R1 genome. Further analysis of these and other putative promoters was performed by Northern hybridization and primer extension experiments. In contrast to previous reports, the -10 and -35 regions of these promoters resembled the sigma 70 consensus sequence of E. coli.

* Corresponding author. Mailing address: Dept. of Chemical Engineering, Box 351750, University of Washington, Seattle, WA 98195-1750. Phone: (206) 616 5282. Fax: (206) 616 5721. E-mail: lidstrom{at}u.washington.edu.

dagger Present address: Department of Molecular Microbiology, Vrije Universiteit Amsterdam, 1081 HV Amsterdam, The Netherlands.

Journal of Bacteriology, May 2001, p. 3169-3175, Vol. 183, No. 10
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.10.3169-3175.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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