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.
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Acknowledgements
We gratefully thank Dr. Ko Kato (Nara Institute of Science and Technology) for valuable discussions and critical reading of the manuscript.
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Koda, A., Bogaki, T., Minetoki, T. et al. 5′ Untranslated region of the Hsp12 gene contributes to efficient translation in Aspergillus oryzae . Appl Microbiol Biotechnol 70, 333–336 (2006). https://doi.org/10.1007/s00253-005-0083-x
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DOI: https://doi.org/10.1007/s00253-005-0083-x