Molecular characterization and physiological function of plant nucleoside transporters

核苷在人类、动物、真菌和原生生物中的转运在过去已经得到了广泛的研究。我们现在已经确定了第一个植物核苷转运蛋白ENT 1从拟南芥的功能水平。明天早上thaliana基因组8种不同的基因编码推定的核苷转运蛋白。我们的目的是确定它们的生化特性并分析相应的生理功能。为此，将cDNA克隆并在异源系统（例如面包酵母或爪蟾卵母细胞）中表达，以确定转运特征。使用GFP融合蛋白，这将是瞬时表达在原生质体中，我们要分析的ENT亚型在细胞内的定位。将通过产生具有启动子-GUS融合体的转基因植物来分析各种ENT启动子在不同组织中以及在植物发育期间的活性。最后，第一次敲除ENT转运蛋白的植物的可用性提供了对表型、发育和差异表达蛋白的相应变化的了解。

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.