Regulatory mechanism of expression of plant thiol-endopeptidase genes

植物硫醇内肽酶基因表达调控机制

• 批准号: 03454011
• 负责人: MINAMIKAWA Takao
• 金额: $ 4.16万
• 依托单位: Tokyo Metropolitan 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-03454011/
• 关键词: Endopeptidase; Legume plant; Promoter (gene); Seed storage protein; Transgenic plant; Gene expression; Seed germination; Protease; エンドペプチダ-ゼ; αーアミラ-ゼ; 種子形成; マメ科種子; 塩基配列; 遺伝子導入

A cysteine endopeptidase (SH-EP) is involved in the degradation of storage globulin in cotyledons of germinating seeds of Vigna mungo seeds. Using SH-EP cDNA as a probe, we isolated two overlapping genomic clones for SH-EP from a V. mungo genomic library. The results of Southern blot analysis of the nuclear DNA suggested that there is a single gene for SH-EP. The site of initiation of transcription was located 31 bp upstream from the initiation codon. Protein immunoblotting showed that, in addition to the presence of the protein in cotyledons of germinating seeds, levels of a protein that reacted with an antiserum against SH-EP increased in pods of maturing fruits of V. mungo, with very low levels being present in other organs of the seedlings. The change in levels of SH-EP mRNA in cotyledons of germinating seeds followed a time course similar to that of alpha-amylase mRNA. Levels of both enzymes, as well as levels of their mRNAs, were also examined in cotyledons of seedlings of three other legumes by protein immunoblotting and RNA blot hybridization, respectively.

半胱氨酸内肽酶（SH-EP）参与绿豆种子萌发过程中子叶贮藏球蛋白的降解。以SH-EP cDNA为探针，从蒙戈弧菌基因组文库中分离到两个重叠的SH-EP基因组克隆。Southern杂交结果表明，SH-EP基因为单基因。转录起始位点位于起始密码子上游31 bp处。蛋白质免疫印迹表明，除了存在于萌发种子的子叶中的蛋白质外，与抗SH-EP的抗血清反应的蛋白质的水平在成熟果实的豆荚中增加，在幼苗的其他器官中存在非常低的水平。萌发种子子叶中SH-EP mRNA水平的变化遵循与α-淀粉酶mRNA相似的时间过程。这两种酶的水平，以及它们的mRNA的水平，也检查了其他三种豆科植物的幼苗子叶蛋白质免疫印迹和RNA印迹杂交，分别。

项目成果

Y.Ogushi,T.Tanaka,D.Yamauchi,T.Minamikawa: "Nucleotide sequence of a gene for an endopeptidase (EP-C1) from Phaseolus vulgaris." Plant Mol.Biol. 19. 705-706 (1992)

Y.Ogushi、T.Tanaka、D.Yamauchi、T.Minamikawa：“菜豆内肽酶 (EP-C1) 基因的核苷酸序列。”

Minamikawa,T.Yamauchi,D.,Wada,S.and Takeuchi,H.: "Expression of αーamylase in Phaalas vulgaris and Vigna mungo plants" Plant Cell Physiol.33. (1992)

Minamikawa, T. Yamauchi, D.、Wada, S. 和 Takeuchi, H.：“α-淀粉酶在 Phaalas vulgaris 和 Vigna mungo 植物中的表达”Plant Cell Physiol 33。（1992）

Ogushi, T., T.Tanaka, D. Yamauchi and T. Minamikawa: "Nucleotide sequence of a gene for an endopeptidase (EP-C1) from Phaseolus vulgaris." Plant Mol. Biol.19. 705-706 (1992)

Ogushi, T.、T.Tanaka、D. Yamauchi 和 T. Minamikawa：“菜豆内肽酶 (EP-C1) 基因的核苷酸序列。”

T.Tanaka,D.Yamauchi,T.Minamikawa: "Nucleotide sequence of cNDA for an endopeptidase (EP-C1) from pods of maturing Phaseolus Vulgaris fruits." Plant Mol.Biol.16. 1083-1084 (1991)

T.Tanaka、D.Yamauchi、T.Minamikawa：“成熟菜豆果实豆荚中肽链内切酶 (EP-C1) 的 cNDA 核苷酸序列。”

T.Tanaka,T.Minamikawa,D.Yamauchi,Y.Ogushi: "Expression of an erdopeptidase (EP-C1) in Phaseolus vulgaris plants." Plant Physiol.101. (1993)

T.Tanaka、T.Minamikawa、D.Yamauchi、Y.Ogushi：“erdopeptidase (EP-C1) 在菜豆植物中的表达。”