Deletion analysis of the superoxide dismutase (sodM) promoter from Aspergillus oryzae.
The manganese superoxide dismutase gene (sodM) is very highly expressed in Aspergillus oryzae. To elucidate the basis for this high-level expression, deletion analysis of the promoter was undertaken using beta-glucuronidase (GUS) as a reporter. Deletion of a 63-bp sequence from -200 to -138 in the 1,038-bp sodM promoter caused a drastic decrease in GUS activity. In addition, an electrophoretic gel mobility shift assay (EMSA) implicated a 30-bp element from -209 to -178 containing cis-element(s) in the high-level expression. The results of fine structure deletion analysis of this region were consistent with the EMSA results. To confirm these findings, we constructed enhanced sodM promoters by incorporating tandem repeats of this region, which resulted in an approximate twofold increase in expression relative to the native sodM promoter.
Hisada
Hiromoto
H
Research Institute, Gekkeikan Sake Co. Ltd. 101 Shimotoba-koyanagi-cho, Fushimi-ku, Kyoto, 612-8361, Japan. hisada@gekkeikan.co.jp
Sano
Motoaki
M
Ishida
Hiroki
H
Hata
Yoji
Y
Abe
Yasuhisa
Y
Machida
Masayuki
M
D001236:Aspergillus oryzae / Q000201:enzymology* / Q000235:genetics; D017353:Gene Deletion; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013482:Superoxide Dismutase / Q000235:genetics* / Q000378:metabolism*
D013482:Superoxide Dismutase