한빛사 논문
Eunjin Byeona,#, Min-Sub Kima,#, Duck-Hyun Kima, Yoseop Leea, Haksoo Jeonga, Jin-Sol Leea, Sung-Ah Hongb, Jun Chul Parkc, Hye-Min Kangd, Alaa El-Din H. Sayede, Yasuhiko Katf, Sangsu Baeb, Hajime Watanabef, Young Hwan Leea,*, Jae-Seong Leea,*
aDepartment of Biological Sciences, College of Science, Sungkyunkwan University, Suwon 16419, South Korea
bDepartment of Chemistry, College of Nature Sciences, Hanyang University, Seoul 04763, South Korea
cDépartement des Sciences, Université Sainte-Anne, Church Point, NS B0W 1M0, Canada
dMarine Biotechnology Research Center, Korea Institute of Ocean Science and Technology, Busan 49111, South Korea
eDepartment of Zoology, Faculty of Sciences, Assiut University, Assiut 71516, Egypt
fDepartment of Biotechnology, Graduate School of Engineering, Osaka University, Osaka 565-0871, Japan
#These authors contributed equally to this work.
*Corresponding author.
Abstract
The water flea Daphnia magna is a small freshwater planktonic animal in the Cladocera. In this study, we assembled the genome of the D. magna NIES strain, which is widely used for gene targeting but has no reported genome. We used the long-read sequenced data of the Oxford nanopore sequencing tool for assembly. Using 3,231 genetic markers, the draft genome of the D. magna NIES strain was built into ten linkage groups (LGs) with 483 unanchored contigs, comprising a genome size of 173.47 Mb. The N50 value of the genome was 12.54 Mb and the benchmarking universal single-copy ortholog value was 98.8%. Repeat elements in the D. magna NIES genome were 40.8%, which was larger than other Daphnia spp. In the D. magna NIES genome, 15,684 genes were functionally annotated. To assess the genome of the D. magna NIES strain for CRISPR/Cas9 gene targeting, we selected glutathione S-transferase omega 2 (GST-O2), which is an important gene for the biotransformation of arsenic in aquatic organisms, and targeted it with an efficient make-up (25.0%) of mutant lines. In addition, we measured reactive oxygen species and antioxidant enzymatic activity between wild type and a mutant of the GST-O2 targeted D. magna NIES strain in response to arsenic. In this study, we present the genome of the D. magna NIES strain using GST-O2 as an example of gene targeting, which will contribute to the construction of deletion mutants by CRISPR/Cas9 technology.
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