Stable transformation of cultured cells of the liverwortMarchantia polymorpha by particle bombardment |
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| Authors: | Kohei Irifune Kanji Ono Misa Takahashi Hideko Murakami Hiromichi Morikawa |
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| Affiliation: | (1) Graduate Department of Gene Sciences, Faculty of Science, Hiroshima University, Kagamiyama, 739 Hagashi-Hiroshima, Japan;(2) Department of Biological Sciences, Faculty of Science, Kumamoto University, Kurokami, 860 Kumamoto, Japan |
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| Abstract: | Suspension-cultured cells (A-18 line) of the liverwortMarchanta polymorpha were bombarded by a pneumatic particle gun with plasmid pCH harbouring the hygromycin phosphotransferase (HPT) gene (hpt) under the control of the cauliflower mosaic virus (CaMV) 35 S promoter and the nopaline synthase polyadenylation region. Nine weeks after bombardments, 128 hygromycin-resistant calluses were obtained from an approximate total of 7×106 cells. Ten cell lines chosen randomly were analysed further. Southern blot analysis showed that all of the ten lines contain thehpt gene in the genome, demonstrating that these lines are transformants. An HPT enzyme activity assay confirmed the expression of the gene in all of the transformant lines. |
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| Keywords: | cultured liverwort cells hygromycin phosphotransferase (HPT) Marchantia polymorpha particle bombardment stable transformation |
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