| 研究実績の概要 |
During the first year of the Kakenhi project, we finished the sequencing of the genome of our bobtail squid model E. berryi using a combination of short and long reads (pacbio) as well as HiC scaffolding. We obtained a 5.91Gb assembly with 46 large chromosomes consistent with chromosome number in other members of the clade Decapoda. The genome was thoroughly annotated, 57259 genes were identified that were subsequently used for gene family reconstruction.Key development gene families (Pax, Wnt, Homeodomain) were analysed using phylogenetic reconstruction to determine the orthodoxy relationships. Our genome assembly harbours a complete Hox cluster spanning over 17Mb which confirms and expands previous observation in other cephalopod species and suggests unusual principles of Hox genes regulation.
We performed preliminary assays of ATAC-seq and RNA-seq for embryonic stages and organs, which is an important resource for the project. During the season 2020, we obtained many embryos and successfully dissected limb buds as planned in the grant. Some samples were preserved for RNA-seq and with others, we tested ATAC-seq experiments using the omni-atac protocol. We managed to obtain ATAC-seq with a good size profile using paired limb pairs (right and left size). There samples are due for sequencing. In parallel, efforts have been made in the lab to establish antibody staining and fluorescent imaging. We are quite confident about the next steps of the project.
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