Studies on the Suppression of Allergenic Proteins in Rice Seeds by Genetic Engineering

基因工程抑制水稻种子致敏蛋白的研究

• 批准号: 03454068
• 负责人: NAKAMURA Ryo
• 金额: $ 3.33万
• 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 (B)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-03454068/
• 关键词: Rice Allergenic Protein; Anti-Sense RNA; Allergenic Protein Promoter; Transgenic Plant; Rice Seeds; Genetic Engineering Study; シスエレメント; 転写調節領域; ゲノムDNA

A genomic clone containing two genes of rice allergenic proteins (RA) was isolated from a rice genomic DNA library. For each RA gene putative TATA and CAAT boxes existed about 50 and 150bp upstream of the transcription initiation site, respectively. Southern blotting analysis also showed that that the allergenic proteins were encoded by a multigene family containing at least four members. To suppress expression of the RA gene during rice seed ripening by anti-sense RNA, anti-sense RA genes were constructed with some plant gene promoters including the RA promoter, and introduced into rice protoplast by electroporation. Transformed cells were selected and grown to maturity. Rice seeds harvested from some of such transgenic plants contained reduced amount of RA, suggesting that the anti-sense RNA method was effective to suppress the RA gene expression in ripening rice seeds.

从水稻基因组DNA文库中分离到一个含有两个水稻过敏蛋白(RA)基因的基因组克隆。对于每个RA基因，在转录起始点上游分别存在约50bp的TATA盒和150bp的CAAT盒。Southern blotting分析还表明，致敏蛋白由一个至少包含四个成员的多基因家族编码。为了通过反义RNA抑制水稻种子成熟过程中RA基因的表达，利用包括RA启动子在内的植物基因启动子构建了反义RA基因，并通过电穿孔的方法将其导入水稻原生质体。筛选转化细胞，培养至成熟。从部分转基因植株中获得的水稻种子含有较少的RA，这表明反义RNA方法可以有效地抑制成熟水稻种子中RA基因的表达。

项目成果

T.Adachi,et al.: "Gene structure and expression of rice seed allergenic proteins belonging to the α-alylase/trypsin inhibitor family." Plant Mol.Biol. 21. 239-248 (1993)

T.Adachi 等人：“属于 α-淀粉酶/胰蛋白酶抑制剂家族的水稻种子过敏蛋白的基因结构和表达。植物分子生物学”21. 239-248 (1993)

T. Matsuda, et al.: "Immunochemical Studies on Rice Allergenic Proteins." Agric. Biol. Chem.55. 509-513 (1991)

T. Matsuda 等人：“大米过敏蛋白的免疫化学研究”。

松田 幹: "Immunochemical Studies on Rice Allergenic Proteins." Agric.Biol.Chem.55. 509-513 (1991)

Miki Matsuda：“大米致敏蛋白的免疫化学研究”。Agric.Biol.Chem.509-513（1991）。

中村 良: "食品分析学(アレルゲン成分の解析)" 文永堂出版, 203-218 (1991)

中村亮：《食品分析（过敏原成分分析）》文内道出版社，203-218（1991）

A.Urisu,et al.: "16-Kilodalton Rice Protein Is One of the Major Allergens in Rice Grain Extract and Responsible for Cross-Allergenicity between Cereal Grains in the Poaceae Family." Int.Arch.Allergy Appl.Immunol.96. 244-252 (1991)

A.Urisu 等人：“16 千道尔顿大米蛋白是大米提取物中的主要过敏原之一，是禾本科谷物之间交叉过敏性的原因。”