NDLI: A further study on chromosome minimization by protoplast fusion in Aspergillus oryzae

Content Provider	Springer Nature Link
Author	Hara, Seiichi Jin, Feng Jie Takahashi, Tadashi Koyama, Yasuji
Copyright Year	2011
Abstract	Our goal in this work was to develop a method to minimize the chromosomes of Aspergillus oryzae, to arrive at a deeper understanding of essential gene functions that will help create more efficient industrially useful strains. In a previous study, we successfully constructed a highly reduced chromosome 7 using multiple large-scale chromosomal deletions (Jin et al. in Mol Genet Genomics 283:1–12, 2010). Here, we have created a further reduced chromosome A. oryzae mutant harboring a reduced chromosome 7 and a reduced chromosome 8 both of which contain a large number of non-syntenic blocks. These are the smallest A. oryzae chromosomes that have been reported. Protoplast fusion between the two distinct chromosome-reduced mutants produced a vigorous and stable fusant which was isolated. PCR and flow cytometry confirmed that two kinds of nuclei, derived from the parent strains, existed in this fusant and that the chromosome DNA per nucleus was doubled, suggesting that the fusant was a heterozygous diploid strain. By treating the cell with 1 μg/ml benomyl, cell nuclei haploidization was induced in the stable diploid strain. Array comparative genomic hybridization and pulsed-field gel electrophoresis confirmed that the reduced chromosomes 7 and 8 co-existed in the haploid fusant and that no other chromosomal modifications had occurred. This method provides a useful tool for chromosome engineering in A. oryzae to construct an industry-useful strain.
Starting Page	177
Ending Page	187
Page Count	11
File Format	PDF
ISSN	16174615
Journal	Molecular and General Genetics MGG
Volume Number	287
Issue Number	2
e-ISSN	16174623
Language	English
Publisher	Springer-Verlag
Publisher Date	2011-12-30
Publisher Place	Berlin/Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Aspergillus oryzae Mutant Fusants Chromosome engineering Microbial Genetics and Genomics Animal Genetics and Genomics Biochemistry Cell Biology Plant Genetics & Genomics
Content Type	Text
Resource Type	Article
Subject	Genetics Medicine Molecular Biology

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