Introduction of a long synthetic repetitive DNA sequence into cultured tobacco cells
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- Ohzeki Junichirou
- Laboratory of Chromosome Engineering, Department of Frontier Research and Development, Kazusa DNA Research Institute
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- Kugou Kazuto
- Laboratory of Chromosome Engineering, Department of Frontier Research and Development, Kazusa DNA Research Institute
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- Otake Koichiro
- Laboratory of Chromosome Engineering, Department of Frontier Research and Development, Kazusa DNA Research Institute
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- Okazaki Koei
- Laboratory of Chromosome Engineering, Department of Frontier Research and Development, Kazusa DNA Research Institute
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- Takahashi Seiji
- Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku University
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- Shibata Daisuke
- Laboratory of Chromosome Engineering, Department of Frontier Research and Development, Kazusa DNA Research Institute
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- Masumoto Hiroshi
- Laboratory of Chromosome Engineering, Department of Frontier Research and Development, Kazusa DNA Research Institute
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抄録
<p>Genome information has been accumulated for many species, and these genes and regulatory sequences are expected to be applied in plants by enhancing or creating new metabolic pathways. We hypothesized that manipulating a long array of repetitive sequences using tethered chromatin modulators would be effective for robust regulation of gene expression in close proximity to the arrays. This approach is based on a human artificial chromosome made of long synthetic repetitive DNA sequences in which we manipulated the chromatin by tethering the modifiers. However, a method for introducing long repetitive DNA sequences into plants has not yet been established. Therefore, we constructed a bacterial artificial chromosome-based binary vector in Escherichia coli cells to generate a construct in which a cassette of marker genes was inserted into 60-kb synthetic human centromeric repetitive DNA. The binary vector was then transferred to Agrobacterium cells and its stable maintenance confirmed. Next, using Agrobacterium-mediated genetic transformation, this construct was successfully introduced into the genome of cultured tobacco BY-2 cells to obtain a large number of stable one-copy strains. ChIP analysis of obtained BY-2 cell lines revealed that the introduced synthetic repetitive DNA has moderate chromatin modification levels with lower heterochromatin (H3K9me2) or euchromatin (H3K4me3) modifications compared to the host centromeric repetitive DNA or an active Tub6 gene, respectively. Such a synthetic DNA sequence with moderate chromatin modification levels is expected to facilitate manipulation of the chromatin structure to either open or closed.</p>
収録刊行物
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- Plant Biotechnology
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Plant Biotechnology 39 (2), 101-110, 2022-06-25
日本植物バイオテクノロジー学会
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詳細情報 詳細情報について
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- CRID
- 1390855511130713984
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- NII論文ID
- 130008161265
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- NII書誌ID
- AA11250821
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- ISSN
- 13476114
- 13424580
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- NDL書誌ID
- 032272046
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- 本文言語コード
- en
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- データソース種別
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- JaLC
- NDL
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- KAKEN
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- 使用不可