Expression of foreign proteins in rice seeds with rice protein gene promoters.

具有水稻蛋白基因启动子的水稻种子中外源蛋白的表达。

• 批准号: 05660134
• 负责人: MATSUDA Tsukasa
• 金额: $ 1.34万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05660134/
• 关键词: rice protein; transgenic rice plant; gene expression; lactoferrin; 形質転換イネ; イネ種子; コメアルブミン遺伝子; GUS融合遺伝子

Fusion genes of rice protein gene promoter and GUS gene were constructed and introduced in rice protoplast by electroporation. Transformed rice plants were regenerated, and expression of the reporter GUS gene was assayd. The GUS activity was specifically detected in seeds of the transgenic rice plants. It was demonstrated that the gluterin gene promoter and the branching enzyme gene promoter were suitable for the expression of foreign genes in seeds of transgenic rice plants.Milk lactoferrin was selected as a model protein to be expressed in seeds of transgenic rice plants, because lactoferrin is a protein in milk commonly consumed as a food and the protein and its N-terminal peptide have anti-bacterial activity. Oligonucleotides encoding the lactoferrin N-terminal peptide chemically synthesized, and double stranded DNA fragment was inserted in a plasmid pUC and E.coli was transformed with the plasmid. The cDNA encoding the lactoferrin was thus obtained. The cDNA was ligated with the above described promoter DNA fragments and were introduced in the rice protoplast. The trangenic rice plants are now regenerating.

构建水稻蛋白基因启动子与GUS基因的融合基因，并通过电穿孔导入水稻原生质体。再生转化的水稻植株，并测定报告GUS基因的表达。在转基因水稻植物的种子中专门检测到了 GUS 活性。证明谷蛋白基因启动子和分支酶基因启动子适合外源基因在转基因水稻种子中的表达。选择牛奶乳铁蛋白作为转基因水稻种子中表达的模型蛋白，因为乳铁蛋白是牛奶中常见的食品蛋白质，且该蛋白质及其N端肽具有抗菌活性。将化学合成的编码乳铁蛋白N端肽的寡核苷酸和双链DNA片段插入质粒pUC中，并用该质粒转化大肠杆菌。由此获得编码乳铁蛋白的cDNA。将cDNA与上述启动子DNA片段连接并引入水稻原生质体中。转基因水稻现在正在再生。

项目成果

Matuda,T.: "Isolation and properties of concanvalian A-like lectin in sword bean." Postharvest biochemistry of plant food-materials in the tropics.17-24 (1994)

Matuda,T.：“刀豆中伴生 A 样凝集素的分离和特性。”

Matsuda T.and Nakamura R.: "Molecular structure and immunological properties of food allergens." Trends food Sci.Tech.4. 289-293 (1993)

Matsuda T. 和 Nakamura R.：“食物过敏原的分子结构和免疫学特性。”

Matsuda, T.and Nakamura, R.: "Molecular structure and immunological properties of food allergens." Trends Food Sci.Tech.4. 289-293 (1993)

Matsuda, T. 和 Nakamura, R.：“食物过敏原的分子结构和免疫学特性。”

Adachi, T., Matsuda, T., Nakamura, R., Alvarez A.M.and Garcia, V.V.: "Allergenic proteins in rice." Japan Scientific Society Press. 3-15 (1994)

Adachi, T.、Matsuda, T.、Nakamura, R.、Alvarez A.M. 和 Garcia, V.V.：“大米中的致敏蛋白”。

Matsuda, T., Kato, H., Takeuchi, S., Nakamura, R., Rodriguez, F.M.and Mendoza, E.M.T.: "Postharvest biochemistry of plant food-materials in the tropics." Japan Scientific Society Press. 17-24 (1994)

Matsuda, T.、Kato, H.、Takeuchi, S.、Nakamura, R.、Rodriguez, F.M. 和 Mendoza, E.M.T.：“热带植物食品材料的采后生物化学”。