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Introduction of a plasmid and a protein into bovine and swine cells by water-in-oil droplet electroporation
http://hdl.handle.net/10659/00007046
http://hdl.handle.net/10659/000070466a65e79b-6a1e-4926-9220-9b851581e693
名前 / ファイル | ライセンス | アクション |
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R-2021-210_kitamura.pdf (2.7 MB)
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Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2022-02-21 | |||||
タイトル | ||||||
タイトル | Introduction of a plasmid and a protein into bovine and swine cells by water-in-oil droplet electroporation | |||||
言語 | ||||||
言語 | eng | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | bovine cell | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | electroporation | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | sperm | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | swine cell | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | bovine cell | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | electroporation | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | sperm | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | swine cell | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
資源タイプ | journal article | |||||
著者 |
ISHINO, Takeshi
× ISHINO, Takeshi× KURITA, Hirofumi× KIRISAWA, Rikio× SHIMAMOTO, Yoshinori× NUMANO, Rika× 北村, 浩 |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | Instrument cost is a major problem for the transduction of DNA fragments and proteins into cells. Water-in-oil droplet electroporation (droplet-EP) was recently invented as a low-cost and effective method for the transfection of plasmids into cultured human cells. We here applied droplet-EP to livestock animal cells. Although it is difficult to transfect plasmids into bovine fibroblasts using conventional lipofection methods, droplet-EP enabled us to introduce an enhanced green fluorescent protein (EGFP)-expressing plasmid into bovine earlobe fibroblasts. The optimal transfection condition was 3.0 kV, which allowed 19.1% of the cells to be transfected. For swine earlobe fibroblasts, the maximum transfection efficacy was 14.0% at 4.0 kV. After transfection with droplet-EP, 69.1% of bovine and 76.5% of swine cells were viable. Furthermore, droplet-EP successfully transduced Escherichia coli recombinant EGFP into frozen-thawed bovine sperm at 1.5 kV. Flow cytometry analysis revealed that 71.5% of spermatozoa exhibited green fluorescence after transfection. Overall, droplet-EP is suitable for the transfection of plasmids and proteins into cultured livestock animal cells. | |||||
書誌情報 |
Journal of Veterinary Medical Science 巻 82, 号 1, p. 14-22, 発行日 2020-01 |
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ISBN | ||||||
識別子タイプ | ISBN | |||||
関連識別子 | 0916-7250 | |||||
DOI | ||||||
関連タイプ | isIdenticalTo | |||||
識別子タイプ | DOI | |||||
関連識別子 | https://doi.org/10.1292/jvms.19-0475 | |||||
権利(URI) | ||||||
権利情報 | https://creativecommons.org/licenses/by-nc-nd/4.0/deed.ja | https://creativecommons.org/licenses/by-nc-nd/4.0/deed.ja | |||||
著者版フラグ | ||||||
出版タイプ | VoR | |||||
出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 | |||||
出版者 | ||||||
出版者 | 日本獣医学会 | |||||
資源タイプ | ||||||
内容記述タイプ | Other | |||||
内容記述 | Article |