Stable nuclear transformation of the diatom Chaetoceros sp. |
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| Authors: | Arisa Miyagawa‐Yamaguchi Takuma Okami Nozomu Kira Haruo Yamaguchi Kouhei Ohnishi Masao Adachi |
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| Institution: | 1. Kochi System Glycobiology Center, Kochi Medical School;2. Laboratory of Aquatic Environmental Science, Faculty of Agriculture;3. Research Institute of Molecular Genetics, Kochi University, Kochi, Japan |
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| Abstract: | A nuclear transformation system for the centric diatom Chaetoceros sp. has been established using two plasmids pTpfcp/nat and pTpNR/green fluorescent protein (GFP) that had been used for Thalassiosira pseudonana transformation. These contain the nourseothricin resistance gene (nat) with the fucoxanthin chlorophyll a/c binding protein (fcp) promoter/terminator from T. pseudonana and the enhanced green fluorescent protein gene (egfp), with the nitrate reductase (NR) promoter/terminator from T. pseudonana, respectively. Transformants were recovered in the presence of the antibiotic nourseothricin. One to four copies of both nat and egfp genes were integrated into genomic DNA of the transformants. Transformation efficiency was 1.5–6.0 transformants per 108 cells. This work is the first report of stable genetic transformation of Chaetoceros, which is important as not only a constituent member of marine ecosystem but also feed for aquaculture. |
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| Keywords: | Chaetoceros diatom transformation |
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