Bioscience, Biotechnology, and Biochemistry
Online ISSN : 1347-6947
Print ISSN : 0916-8451
Biochemistry & Molecular Biology Regular Papers
Gene Silencing by RNA Interference in the Koji Mold Aspergillus oryzae
Osamu YAMADARyoko IKEDAYuka OHKITARisa HAYASHIKazutoshi SAKAMOTOOsamu AKITA
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JOURNAL FREE ACCESS

2007 Volume 71 Issue 1 Pages 138-144

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Abstract

We found the orthologous genes required for RNA interference (RNAi) in the Aspergillus oryzae genome database, and constructed a set of tools for gene silencing using RNAi in A. oryzae. This system utilizes compatible restriction enzyme sites so that only a single target gene fragment is required to create the hairpin RNA cassette. For ease of handling, we also separated the construction of the hairpin RNA cassette for the target gene from its subsequent introduction into the expression vector. Using the brlA gene as a target for RNAi, we detected decreased mRNA levels and a delayed conidiation phenotype in the transformants. Furthermore, even though A. oryzae possesses three copies of the α-amylase gene, a single copy of an α-amylase RNAi construct was sufficient to downregulate the mRNA levels and decrease the enzymatic activity to 10% of control levels. Gene silencing by RNAi should provide a powerful genetic tool for post-genomic studies of the industrially important fungus A. oryzae.

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© 2007 by Japan Society for Bioscience, Biotechnology, and Agrochemistry
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