Applied Genetics and Molecular Biotechnology

Applied Microbiology and Biotechnology

, 70:333

First online:

5′ Untranslated region of the Hsp12 gene contributes to efficient translation in Aspergillus oryzae

  • Akio KodaAffiliated withGeneral Research Laboratory, Ozeki Corp. Email author 
  • , Takayuki BogakiAffiliated withGeneral Research Laboratory, Ozeki Corp.
  • , Toshitaka MinetokiAffiliated withGeneral Research Laboratory, Ozeki Corp.
  • , Masato HirotsuneAffiliated withGeneral Research Laboratory, Ozeki Corp.

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Abstract

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°C). Moreover, at high temperature (37°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.