Development of a promoter shutoff system in Aspergillus oryzae using a sorbitol-sensitive promoter. Issue 9 (1st September 2016)

• Record Type: Journal Article
• Authors: Oda, Ken; Terado, Shiho; Toyoura, Rieko; Fukuda, Hisashi; Kawauchi, Moriyuki; Iwashita, Kazuhiro
• Is Part Of: Bioscience, biotechnology, and biochemistry. Volume 80:Issue 9(2016)
• Page Start: 1792
• Page End: 1801
• Publication Date: 2016-09-01
• Subjects: Aspergillus oryzae -- promoter shutoff -- essential gene analysis -- sorbitol metabolism; Biotechnology -- Periodicals; Biochemistry -- Periodicals; 660.6
• DOI: 10.1080/09168451.2016.1189313
• Languages: English
• ISSNs: 0916-8451

Abstract

Abstract: Promoter shutoff is a general method for analyzing essential genes, but in the fungus Aspergillus oryzae, no tightly repressed promoters have been reported. To overcome the current limitations of conditional promoters, we examined sorbitol- and galactose-responsive genes using microarrays to identify regulatable genes with only minor physiological and genetic effects. We identified two sorbitol-induced genes (designated as sorA and sorB ), cloned their promoters, and built a regulated egfp and brlA expression system. Growth medium-dependent enhanced green fluorescence protein (EGFP) fluorescence and conidiation were confirmed for egfp and brlA under the control of their respective promoters. We also used this shutoff system to regulate the essential rhoA, which demonstrated the expected growth inhibition under repressed growth conditions. Our new sorbitol promoter shutoff system developed can serve as a valuable new tool for essential gene analyses of filamentous fungi. Graphical abstract: : A sorbitol-responsive promoter shutoff system was evaluated for egfp and essential rhoA expression controlled with P sorA and P sorB in A. oryzae .