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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,
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
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.

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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