Molecular characterization and physiological function of plant nucleoside transporters

植物核苷转运蛋白的分子特征和生理功能

• 批准号: 5386750
• 负责人: Dr. Torsten Möhlmann
• 金额: --
• 依托单位: Fachgebiet Pflanzenphysiologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2002
• 资助国家: 德国
• 起止时间: 2001-12-31 至 2007-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5386750?language=en
• 关键词: Molecular; characterization; physiological; function; plant

Nucleoside transport across human, animal, fungi and protists has already been intensively characterised in the past. We have now identified the first plant nucleoside transporter ENT1 from Arabidopsis thaliana on a functional level. In the A. thaliana genome 8 different genes encode putative nucleoside transporters. Our aim is to determine their biochemical properties and analyse the corresponding physiological function. To do this the cDNA's will be cloned and expressed in heterologous systems (e.g. bakers yeast or xenopus oocytes) to determine the transport characteristics. Using GFP fusion proteins, which will be transiently expressed in protoplasts, we want to analyse the localisation of ENT isoforms within the cell. The activity of the various ENT promoters in different tissues, and during plant development will be analysed by generating transgenic plants with promoter-GUS fusions. Finally, the availability of knock out plants for ENT transporters for the first time provides insight in corresponding changes in phenotypes, development, and differentially expressed proteins.

核苷在人类、动物、真菌和原生生物之间的运输在过去已经得到了广泛的描述。我们现在已经在功能水平上从拟南芥中鉴定出第一个植物核苷转运蛋白ENT1。在拟南芥基因组中，有8个不同的基因编码假定的核苷转运体。我们的目的是测定它们的生化性质，并分析相应的生理功能。为此，将克隆并在异源系统(例如面包师、酵母或非洲爪哇卵母细胞)中表达这些基因，以确定其转运特性。利用将在原生质体中瞬时表达的GFP融合蛋白，我们想要分析ENT亚型在细胞内的定位。将通过产生带有启动子-GUS融合的转基因植株来分析各种ENT启动子在不同组织中和植物发育期间的活性。最后，首次获得用于ENT转运蛋白的敲除植物提供了对表型、发育和差异表达蛋白质的相应变化的洞察。