Cloning of a rhodococcal promoter using a transposon for dibenzothiophene biodesulfurization.
The expression of biodesulfurization genes (dsz) in Rhodococcus erythropolis strain KA2-5-1 is repressed by sulfate which is the product of biodesulfurization. The application of a sulfate non-repressible promoter could be effective in enhancing biodesulfurization. A promoter-probe transposon was constructed using the promoterless, red-shifted green fluorescence protein gene (rsgfp). A 340 bp putative promoter element, designated kap1, was isolated from a strain KA2-5-1 recombinant that had shown high fluorescence intensity. The activity of kap1 was not affected by 1 mM sulfate. It gave about a 2-fold greater activity than the 16S ribosomal RNA promoter in R. erythropolis strain KA2-5-1 and is therefore useful for expressing desulfurization genes in rhodococcal strains.
Noda
Ken-ichi
K
Bio-Refining Process Laboratory Technical Cooperation Department, Japan Cooperation Center, Petroleum, 1900 Sodeshi-cho, Shimizu-shi, Shizuoka 424-0037, Japan. nodake@brpl.jccp.or.jp
Watanabe
Kimiko
K
Maruhashi
Kenji
K
D001673:Biodegradation, Environmental; D003001:Cloning, Molecular / Q000379:methods*; D004251:DNA Transposable Elements / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods; D012240:Rhodococcus / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D013045:Species Specificity; D013431:Sulfates / Q000378:metabolism; D013455:Sulfur / Q000378:metabolism*; D013876:Thiophenes / Q000378:metabolism*
D004251:DNA Transposable Elements; D013431:Sulfates; D013876:Thiophenes; D013455:Sulfur; C016366:dibenzothiophene