Preparation of Knockdown Transformants of Unicellular Charophycean Alga, Closterium peracerosum-strigosum-littorale Complex
Jun Abe,
Naoko Hirano,
Ayumi Komiya,
Naho Kanda,
Anri Fujiwara,
Sachie Hori,
Yuki Tsuchikane,
Hiroyuki Sekimoto
Affiliations
Jun Abe
Department of Chemical and Biological Sciences, Faculty of Science, Japan Women’s University, Bunkyo-ku, Tokyo, Japan
Naoko Hirano
Division of Material and Biological Sciences, Graduate School of Science, Japan Women’s University, Bunkyo-ku, Tokyo, Japan
Ayumi Komiya
Division of Material and Biological Sciences, Graduate School of Science, Japan Women’s University, Bunkyo-ku, Tokyo, Japan
Naho Kanda
Division of Material and Biological Sciences, Graduate School of Science, Japan Women’s University, Bunkyo-ku, Tokyo, Japan
Anri Fujiwara
Division of Material and Biological Sciences, Graduate School of Science, Japan Women’s University, Bunkyo-ku, Tokyo, Japan
Sachie Hori
Division of Material and Biological Sciences, Graduate School of Science, Japan Women’s University, Bunkyo-ku, Tokyo, Japan
Yuki Tsuchikane
Department of Chemical and Biological Sciences, Faculty of Science, Japan Women’s University, Bunkyo-ku, Tokyo, Japan
Hiroyuki Sekimoto
Department of Chemical and Biological Sciences, Faculty of Science, Japan Women’s University, Bunkyo-ku, Tokyo, JapanDivision of Material and Biological Sciences, Graduate School of Science, Japan Women’s University, Bunkyo-ku, Tokyo, Japan
To prepare the knockdown transformants of the Closterium peracerosum-strigosum-littorale (C. psl.) complex, particle bombardment was applied with a newly constructed vector (pSA0104) with an endogenous constitutive promoter fused to a DNA fragment corresponding to an antisense strand of a target gene. Using a hygromycin resistance gene (aph7”), hygromycin-resistant colonies were selected. After the second screening, integration of the vector into the genome was checked by PCR and the knockdown effect was evaluated by Western blotting using a specific antibody against the target protein.
