由綠豆(Vigna radiate (L.) Wilczek)胚基因庫分離出一個新型脫水蛋白質(dehydrin)，並命名為VrDhn1。VrDhn1是一個intronless基因且屬於Y2K類型脫水蛋白。利用western blot，顯示VrDhn1大量累積在成熟種子的胚和子葉並且在種子發芽約2天後消失。由反轉錄聚合酶連鎖反應(RT-PCR)及即時聚合酶連鎖反應(real-time PCR)，顯示VrDhn1轉錄子大量表現於經由脫水、高鹽、ABA處理之10天大的綠豆幼苗，卻無法偵測VrDhn1蛋白質的累積。VrDhn1:GFP 融合蛋白(fusion protein)在阿拉伯芥轉殖株生長在一般環境是呈現被降解(degraded)情形，但轉殖株處在脫水的情況下，融合蛋白的表現量比一般生長情況有較穩定存在轉植株內。利用免疫定位(immunolocalization)顯示VrDhn1累積在成熟種子及種子發芽一天後的胚之細胞核及細胞質，且大部分的VrDhn1聚集在核染色質(chromatin)的周圍。Electrophoretic mobility shift assay (EMSA)的分析顯示VrDhn1與DNA片段有不專一性結合，鋅離子及鎳離子會提升VrDhn1與DNA間交互作用(internation)。利用CO-immunoprecipitation實驗顯示綠豆種子內VrDhn1的tyrosine 和 serine胺基酸可被磷酸化(phosphorylation)。VrDhn1 (30.17 kDa)經由膠體過濾法(gel filtration)分析，其分子量為63.1 kDa，推測VrDhn1以二聚物(dimmer)的形式存在。這是第一篇紀錄Y2K類型脫水蛋白質 VrDhn1在種子成熟期間，會進入細胞核並與DNA互相連結的研究。

A novel dehydrin gene (VrDhn1) was isolated from an embryo cDNA library of Vigna radiata (L.) Wilczek (mungbean) variety VC1973A. The intronless VrDhn1 gene encodes a protein belonging to the Y2K-type dehydrin family. VrDhn1 protein accumulated in embryos and cotyledons during seed maturation and disappeared 2 days after seed imbibition (DAI). The expression of VrDhn1 mRNA and accumulation of VrDhn1 protein were at high levels in mature seeds, but neither mRNA nor protein was detected in mungbean vegetative tissues under normal growth conditions. VrDhn1 mRNA level was extremely high in mature seeds and decreased to ~30% at 1 DAI, and was not detectable at ~7 DAI. Tissue dehydration, salinity and exogenous abscisic acid (ABA) markedly induced VrDhn1 transcripts in plants as measured by quantitative real-time reverse transcription-PCR (qRT-PCR). VrDhn1 protein was not detected using immunoblots in seedlings under stress treatments. VrDhn1:GFP fusion protein is degraded in transgenic Arabidopsis under normal condition, but is more stable under dehydration. In mature seeds or 1 DAI seedlings, VrDhn1 proteins were immunolocalized in the nucleus and cytoplasm. VrDhn1 exhibited low affinity of non-specific interaction with DNA using electrophoretic mobility shift assays (EMSA), and the exogenous addition of Zn2+ or Ni2+ stimulated interaction. Phosphorylation occurs on threonine and tyrosine residues of VrDhn1 in mungbean seeds. The His-tagged VrDhn1 (30.17 kDa) protein showed a molecular mass of 63.1 kDa on gel filtration, suggesting a dimer form. This is the first report showing that a Y2K-type VrDhn1 enters the nucleus and interacts with DNA during seed maturation.

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