Promoter cloning in the radioresistant bacterium Deinococcus radiodurans.
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.
Meima
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Department of Chemical Engineering, University of Washington, Seattle, WA 98195, USA.
Rothfuss
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HM
Gewin
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Lidstrom
M E
ME
D050557:Catechol 2,3-Dioxygenase; D003001:Cloning, Molecular; D003470:Culture Media; D049308:Dioxygenases; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D006095:Gram-Positive Cocci / Q000235:genetics* / Q000254:growth & development / Q000528:radiation effects; D007763:Lac Operon / Q000235:genetics / Q000502:physiology; D016296:Mutagenesis; D010105:Oxygenases / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics* / Q000502:physiology; D011836:Radiation Tolerance; D014158:Transcription, Genetic; D014169:Transformation, Bacterial
D003470:Culture Media; D010105:Oxygenases; D049308:Dioxygenases; D050557:Catechol 2,3-Dioxygenase