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Applied and Environmental Microbiology, September 2009, p. 5943-5951, Vol. 75, No. 18
0099-2240/09/$08.00+0     doi:10.1128/AEM.00975-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Identification of a Basic Helix-Loop-Helix-Type Transcription Regulator Gene in Aspergillus oryzae by Systematically Deleting Large Chromosomal Segments{triangledown} ,{dagger}

Feng Jie Jin,1* Tadashi Takahashi,1 Masayuki Machida,2 and Yasuji Koyama1

Noda Institute for Scientific Research, 399 Noda, Noda City 278-0037, Japan,1 Research Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology, Central 6, Higashi 1-1-1, Tsukuba, Ibaraki 305-8566, Japan2

Received 29 April 2009/ Accepted 15 July 2009

We previously developed two methods (loop-out and replacement-type recombination) for generating large-scale chromosomal deletions that can be applied to more effective chromosomal engineering in Aspergillus oryzae. In this study, the replacement-type method is used to systematically delete large chromosomal DNA segments to identify essential and nonessential regions in chromosome 7 (2.93 Mb), which is the smallest A. oryzae chromosome and contains a large number of nonsyntenic blocks. We constructed 12 mutants harboring deletions that spanned 16- to 150-kb segments of chromosome 7 and scored phenotypic changes in the resulting mutants. Among the deletion mutants, strains designated {Delta}5 and {Delta}7 displayed clear phenotypic changes involving growth and conidiation. In particular, the {Delta}5 mutant exhibited vigorous growth and conidiation, potentially beneficial characteristics for certain industrial applications. Further deletion analysis allowed identification of the AO090011000215 gene as the gene responsible for the {Delta}5 mutant phenotype. The AO090011000215 gene was predicted to encode a helix-loop-helix binding protein belonging to the bHLH family of transcription factors. These results illustrate the potential of the approach for identifying novel functional genes.

* Corresponding author. Mailing address: Noda Institute for Scientific Research, 399 Noda, Noda City 278-0037, Japan. Phone: 81-4-7123-5573. Fax: 81-4-7123-5953. E-mail: hkin{at}mail.kikkoman.co.jp

{triangledown} Published ahead of print on 24 July 2009.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, September 2009, p. 5943-5951, Vol. 75, No. 18
0099-2240/09/$08.00+0     doi:10.1128/AEM.00975-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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