The expression analysis of symbiotic nitrogen-carbon

共生氮碳表达分析

• 批准号: 12660061
• 负责人: TAJIMA Shigeyuki
• 金额: $ 2.56万
• 依托单位: Kagawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12660061/
• 关键词: metabolism; transgenic plant; symbiotic nitrogen fixation; cloning; uricase; high level expression; promoter analysis; gel shift assay; 窒素固定; マメ科植物; プリン代謝; 窒素代謝; アラントイン; ウレイド; ウリカーゼ遺伝子; ミヤコグサ

We have cloned and sequenced uricase genes of various legume plants and made a phylogenetic tree. As a result, phylogenetic analysis of uricase sequences from various legumes indicated that uricases of amide- and ureide- transporting legumes forms two distinct groups, suggesting these types of legumes were evolutionary divided.The two uricase genes of soybean were expressed at high level using pET vector. After purification of two uricases, we investigated these uricase characterizations. There was no different Km value against oxygen between them. However, one uricase (UR9) showed lower Km value against uric acid than another uricase (UR2).Gain of function and loss of function experiments suggested that 6 base (GTAATG) motif in the 5' non-coding region of soybean uricase gene was a cis element for nodule uninfected cell specific expression.We produced the transgenic lotus plants for high level expression of PEPC or PPDK to analyze C or N metabolism in nodule.

本研究对多种豆科植物的尿酸酶基因进行了克隆和序列测定，并构建了系统发育树。结果表明，酰胺转运型和酰脲转运型豆科植物的尿酸酶可分为两个不同的类群，表明这两类豆科植物在进化上是不同的。两种尿酸酶纯化后，我们研究了这些尿酸酶的特性。它们对氧的Km值无差异。通过功能获得和功能丧失实验，证明大豆尿酸酶基因5'端非编码区的6个碱基（GTAATG）基序是根瘤非感染细胞特异表达的顺式元件。

项目成果

Takane, K., Tajima, S., Kouchi, H.: "Structural and expression analysis of uricase mRNA from Lotus japonicus"Plant Microbe Interact. 13. 1156-1160 (2000)

Takane, K.、Tajima, S.、Kouchi, H.：“莲花尿酸酶 mRNA 的结构和表达分析”植物微生物相互作用。

Cheng X G,Nomura M,Takane K,Kouchi H Tajima S: "Expression of two uricase (Nodulin-35) genes in a non-ureide type legume Medicago sativa."Plant Cell Physiol.. 41・1. 104-109 (2000)

Cheng X G、Nomura M、Takane K、Kouchi H Tajima S：“非酰脲型豆科植物苜蓿中两种尿酸酶（Nodulin-35）基因的表达。植物细胞生理学.. 104-109（2000）” ）

Tajima S., Takane, K., Nomura, M. and Kouchi, H.: "Symbiotic nitrogen fixation at the late stage of nodule formation in Lotus japonicus and other legume plants"J. Plant Res.. 113. 467-473 (2000)

Tajima S.，Takane，K.，Nomura，M.和Kouchi，H.：“百脉根和其他豆科植物根瘤形成后期的共生固氮”J。

Cheng, X.G: "Expression of two uricase (Nodulin-35) genes in a non-ureide type legume, Medicago sativa"Plant Cell Physiol. 41. 104-109 (2000)

Cheng，X.G：“两种尿酸酶（Nodulin-35）基因在非酰脲型豆科植物苜蓿中的表达”植物细胞生理学。

Shigeyuki Tajima: "Symbiotic nitrogen fixation at the late stage of nodule formation in Lotus japonicus and other legume plants"J. Plant Res.. 113. 467-473 (2000)

Shigeyuki Tajima：“百脉根和其他豆科植物根瘤形成后期的共生固氮”J.