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Stable nuclear transformation of the diatom Chaetoceros sp.
Author(s) -
MiyagawaYamaguchi Arisa,
Okami Takuma,
Kira Nozomu,
Yamaguchi Haruo,
Ohnishi Kouhei,
Adachi Masao
Publication year - 2011
Publication title -
phycological research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.438
H-Index - 44
eISSN - 1440-1835
pISSN - 1322-0829
DOI - 10.1111/j.1440-1835.2011.00607.x
Subject(s) - thalassiosira pseudonana , biology , transformation (genetics) , green fluorescent protein , plasmid , terminator (solar) , chaetoceros , transformation efficiency , nitrate reductase , electroporation , gene , diatom , microbiology and biotechnology , botany , genetics , biochemistry , agrobacterium , nutrient , enzyme , ecology , ionosphere , physics , phytoplankton , astronomy
SUMMARY 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 10 8 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.