Koda Akio, Bogaki Takayuki, Minetoki Toshitaka, Hirotsune Masato
General Research Laboratory, Ozeki Corp., 4-9, Imazu, Dezaike-cho, Nishinomiya-shi, Hyogo, 663-8227, Japan.
Appl Microbiol Biotechnol. 2006 Apr;70(3):333-6. doi: 10.1007/s00253-005-0083-x. Epub 2005 Jul 30.
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.
我们描述了一个5'非翻译区(5'UTR),它能显著提高外源基因在米曲霉中的表达水平。利用一系列5'UTR::GUS(uidA)融合构建体,我们分析了不同5'UTR的嵌合mRNA在不同温度下的翻译效率。我们发现,热休克蛋白基因Hsp12的5'UTR在常温(30℃)下极大地提高了嵌合GUS mRNA的翻译效率。此外,在高温(37℃)下,含有Hsp12 5'UTR的mRNA的翻译效率远高于含有非热休克5'UTR的mRNA,从而使GUS蛋白的表达效率更高(与对照构建体相比,GUS活性高约20倍)。这个5'UTR可与各种强启动子结合使用,以增强外源蛋白在米曲霉中的表达。