Improvement of the Aspergillus oryzae enolase promoter (P-enoA) by the introduction of cis-element repeats.
We constructed a protein expression vector with an improved enoA promoter that harbored 12 tandem repeats of the cis-acting element (region III) of Aspergillus oryzae. The improved promoter yielded reporter beta-glucuronidase (GUS) activity approximately 30-fold of the original promoter. Northern blot analysis confirmed that GUS expression was increased at the transcriptional level. The transformant harboring seven copies of the novel vector showed more than 100,000 U/mg GUS protein, which was approximately 30% of all the cell-free soluble proteins.
Tsuboi
Hirokazu
H
General Research Laboratory, Ozeki Corporation, Hyogo, Japan. hirokazu.tsuboi@ozeki.co.jp
Koda
Akio
A
Toda
Tomomi
T
Minetoki
Toshitaka
T
Hirotsune
Masato
M
Machida
Masayuki
M
D001236:Aspergillus oryzae / Q000201:enzymology / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal; D017930:Genes, Reporter; D005822:Genetic Vectors; D005966:Glucuronidase / Q000378:metabolism; D010751:Phosphopyruvate Hydratase / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D020080:Tandem Repeat Sequences
D005966:Glucuronidase; D010751:Phosphopyruvate Hydratase