DNA transformation into rice protoplasts and the gene expression in the regenerated plants

DNA转化水稻原生质体及其在再生植株中的基因表达

• 批准号: 62480029
• 负责人: HINATA Kokichi
• 金额: $ 4.1万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-62480029/
• 关键词: Rice; Oryza sativa L.; protoplast; electroporation; transgenic plant; APH(3')II gene; GUS gene; APH(3')II遺伝子; トランジエント遺伝子発現

Whole plants of rice (Oryza sativa L.) were derived from protoplasts, which had been electroporated with plasmid DNA possessing a chimeric gene encoding aminoglycoside phosphotransferase II (APH(3')II, conferring resistance to kanamycin and G418) and the cauliflower mosaic virus 35S promoter. Effective conditions of electroporation were determined by monitoring transient expression of the GUS gene (Clontech Lab. Inc.). Transformed calli were selected on the basis of tolerance to the antibiotic, G418. Several regenerated plants possessed functional APH(3')II activity due to the integration of intact foreign DNA into their genome as confirmed by Southern blot analysis.

水稻(Oryza sativa L.)将含有编码氨基糖苷磷酸转移酶II(APH(3‘)II，抗卡那霉素和G418的嵌合基因的质粒DNA和花椰菜花叶病毒35S启动子电转化至原生质体。通过监测GUS基因的瞬时表达(Clontech Lab.Inc.)。根据对抗生素G418的耐受性筛选转化的愈伤组织。Southern印迹分析证实，部分再生植株具有APH(3‘)II活性，这是由于完整的外源DNA整合到其基因组中所致。

项目成果

K.TORIYAMA,et al.: BIO/TECHNOLOGY. 6. 1072-1074 (1988)

K.TORIYAMA 等人：生物/技术。

日向康吉;鳥山欽哉: 育種学雑誌別冊1. 38. (1988)

Yasukichi Hinata；Kinya Toriyama：育种杂志特刊 1. 38. (1988)

Toriyama, K.: "Transgenic rice plants after direct gene transfer into protoplasts." Bio/technology. 6. 1072-1074 (1988)

Toriyama, K.：“直接将基因转移到原生质体后的转基因水稻植物。”

K.Toriyama;et al: BIO/TECHNOLOGY. 6. 1072-1074 (1988)

K.Toriyama；等人：生物/技术。