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Research note: High efficiency transformation of the diatom Phaeodactylum tricornutum with a promoter from the diatom Cylindrotheca fusiformis
Author(s) -
Miyagawa Arisa,
Okami Takuma,
Kira Nozomu,
Yamaguchi Haruo,
Ohnishi Kouhei,
Adachi Masao
Publication year - 2009
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.1440-1835.2009.00531.x
Subject(s) - phaeodactylum tricornutum , biology , diatom , transformation (genetics) , plasmid , gene , microbiology and biotechnology , botany , biochemistry
SUMMARY A high efficiency transformation system was established for the pennate diatom Phaeodactylum tricornutum Bohlin using a plasmid containing fucoxanthin chlorophyll a/c binding protein ( fcp ) promoter/terminator and nitrate reductase ( NR ) promoter/terminator that are derived from the pennate diatom Cylindrotheca fusiformis . The plasmid that contains the zeocin resistance gene ( ble ) with the fcp promoter and enhanced green fluorescent protein gene ( egfp ) with the NR promoter was introduced into P. tricornutum using microparticle bombardment. Transformants (650 ± 58 per 10 8 cells) were obtained. The yield of transformants was between 1.5 and 130 times higher than previously reported P. tricornutum transformation systems. Four to seven copies of the ble gene were integrated into genomic DNA of the transformants. This high efficiency transformation system of P. tricornutum is expected to provide a powerful tool for high‐throughput analysis of gene function using homologous recombination or RNAi.