We found a match
Your institution may have rights to this item. Sign in to continue.
- Title
Identifying the inserted locus of randomly integrated expression plasmids by whole-genome sequencing of Aspergillus strains.
- Authors
Takeda, Aya K.; Udagawa, Hiroaki; Uemura, Yasuo; Takagi, Shinobu
- Abstract
Whole-genome sequencing was conducted on two Aspergillus oryzae strains used for the manufacturing of food enzymes, Acrylaway® and Shearzyme®, with the aim of identifying the inserted locus of randomly integrated expression plasmid and obtaining flanking sequences for safety assessment. Illumina paired-end sequencing was employed, and the obtained reads were mapped to two references: the public genome sequence of Aspergillus oryzae RIB40 and the in-house sequence of the used expression plasmid. Introducing the concept of linking-reads, one locus for each was successfully identified as the integrated site. In the case of Acrylaway®, the obtained sequences suggested that the expression plasmid had been integrated as multiple copies in tandem form. In the case of Shearzyme®, however, information on one edge of the insert was missing, which required extra polymerase chain reaction (PCR) cloning for safety assessment. A 4-kb deletion was detected at the integrated site. There was also evidence of rearrangement occurring in Shearzyme® strain.
- Subjects
PLASMIDS; NUCLEOTIDE sequencing; ASPERGILLUS
- Publication
Bioscience, Biotechnology & Biochemistry, 2018, Vol 82, Issue 11, p1880
- ISSN
0916-8451
- Publication type
Article
- DOI
10.1080/09168451.2018.1506312