Insertion analysis of putative functional elements in the promoter region of the Aspergillus oryzae Taka-amylase A gene (amyB) using a heterologous Aspergillus nidulans amdS-lacZ fusion gene system.
Expression of the Taka-amylase A gene (amyB) of Aspergillus oryzae is induced by starch or maltose. The A. oryzae amyB gene promoter contains three highly conserved sequences, designated Regions I, II, and III, compared with promoter regions of the A. oryzae glaA encoding glucoamylase and the agdA encoding alpha-glucosidase. To identify the function of these sequences within the amyB promoter, various fragments containing conserved sequences in the amyB promoter were introduced into the upstream region of the heterologous A. nidulans amdS gene (encoding acetamidase) fused to the Escherichia coli lacZ gene as a reporter. Introduction of the sequence between -290 to -233 (the number indicates the distance in base pairs from the translation initiation point (+1)) containing Region III significantly increased the expression of the lacZ reporter gene in the presence of maltose. The sequence between -377 to -290 containing Region I also increased the lacZ activity, but its maltose inducibility was less than that of Region III. The sequence between -233 to -181 containing Region II had no effect on the expression. These results indicated that Region III is most likely involved in the maltose induction of the amyB gene expression.
Kanemori
Y
Y
Research Institute of Gekkeikan Sake Co., Ltd., Kyoto, Japan.
Gomi
K
K
Kitamoto
K
K
Kumagai
C
C
Tamura
G
G
D000581:Amidohydrolases / Q000235:genetics; D019677:Artificial Gene Fusion; D001233:Aspergillus nidulans / Q000201:enzymology / Q000235:genetics; D001236:Aspergillus oryzae / Q000201:enzymology* / Q000235:genetics*; D001483:Base Sequence; D017124:Conserved Sequence; D004271:DNA, Fungal / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D005800:Genes, Fungal; D017930:Genes, Reporter; D006026:Glycoside Hydrolases / Q000235:genetics*; D007763:Lac Operon; D008320:Maltose / Q000494:pharmacology; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D014170:Transformation, Genetic; D000516:alpha-Amylases / Q000235:genetics*
D004271:DNA, Fungal; D008320:Maltose; D006026:Glycoside Hydrolases; D000516:alpha-Amylases; C100181:pullulanase; D000581:Amidohydrolases; C021445:acetamidase