5' Untranslated region of the Hsp12 gene contributes to efficient translation in Aspergillus oryzae.
We describe a 5' untranslated region (5'UTR) that dramatically increases the expression level of an exogenous gene in Aspergillus oryzae. Using a series of 5'UTR::GUS (uidA) fusion constructs, we analyzed the translation efficiency of chimeric mRNAs with different 5'UTRs at different temperatures. We found that the 5'UTR of a heat-shock protein gene, Hsp12, greatly enhanced the translation efficiency of the chimeric GUS mRNA at normal temperature (30 degrees C). Moreover, at high temperature (37 degrees C), the translation efficiency of the mRNA containing the Hsp12 5'UTR was far superior to that of mRNAs containing nonheat-shock 5'UTRs, resulting in much more efficient expression of GUS protein (about 20-fold higher GUS activity compared to the control construct). This 5'UTR can be used in combination with various strong promoters to enhance the expression of foreign proteins in A. oryzae.
Koda
Akio
A
General Research Laboratory, Ozeki Corp., 4-9, Imazu, Dezaike-cho, Nishinomiya-shi, Hyogo, 663-8227, Japan. akio.kouda@ozeki.co.jp
Bogaki
Takayuki
T
Minetoki
Toshitaka
T
Hirotsune
Masato
M
D020121:5' Untranslated Regions / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001236:Aspergillus oryzae / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D015966:Gene Expression Regulation, Fungal; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
D020121:5' Untranslated Regions; D006360:Heat-Shock Proteins; D011993:Recombinant Fusion Proteins; D005966:Glucuronidase