臺灣博碩士論文加值系統

蛋白質精胺酸甲基化是常見的轉譯後修飾，將 S-adenosyl-L-methionine （AdoMet）上的甲基集團轉到氮原子上。蛋白質精胺酸甲基轉移酶（PRMT）會調控細胞的訊息傳遞、轉錄、RNA 運輸以及 RNA 剪接等。人類細胞中PRMT1 所催化的精胺酸甲基化反應約佔85 %。PRMT1 缺乏的老鼠胚胎會在 6.5 天的時候死亡。斑馬魚( Danio rerio) 已經成為研究胚胎發育、脊椎動物的遺傳以及人類疾病的重要的模式動物。為了要再更深入了解 prmt1 於胚胎發育的角色，我們以斑馬魚為實驗對象探討 prmt1 於胚胎發育時的功能。斑馬魚的 prmt1 胺基酸序列與人類的相似度為 96 ﹪ 可轉譯出341個胺基酸。取斑馬魚胚胎萃取液經由西方墨點法的分析可證實的確有PRMT1蛋白表現。此外，利用全體原位雜交的方法偵測到 prmt1 自合子期至幼魚早期都有表現。發現 prmt1 在嗅球、眼頂蓋、中腦、小腦、後腦、中後腦交界處以及下巴的咽喉腮弓等部位的細胞上都有明顯的訊號表現。接著以prmt1 以反意 morpholino 將斑馬魚胚胎之 prmt1 抑制，造成了一些 morphant 的缺陷包括：尾巴微彎、體型彎曲、體軸變短、脊椎彎曲、卵黃形狀改變等。以上這些實驗結果顯示了PRMT1對於斑馬魚胚胎發育之影響。

Protein arginine methylation is a widespread post translational modification that transfers methyl groups from S-adenosyl-L
-methionine (AdoMet) to the guanidino nitrogen atoms of arginine residues in the living cells. Protein arginine methyltransferases (PRMT) has been reported to regulate signal transduction, transcription, RNA transport, and RNA splicing. PRMT1 is the predominant arginine methyltransferases in human cell, responsible for at least 85% of all arginine methylation reactions. PRMT1 null mice embryos die at approximately embryonic day 6.5. Zebrafish ( Danio rerio) has recently emerged as an important model organism for studies of embryonic development, vertebrate genetics and human disease. To elucidate the role of PRMT1 during embryogenesis, we use zebrafish as our model system. The zebrafish PRMT1 ORF encodes a 341 amino-acid polypeptide which shares 96% homologies to human PRMT1. Western blot analyses of zebrafish embryo extracts confirmed the expression of PRMT1 protein in zebrafish. During embryonic development, prmt1 transcripts were detected from the zygote period to the early larva stage. Whole-mount in situ hybridization revealed that prmt1 was expressed in most parts of the brain including olfactory bulbs, optic tectum, mesencephalon, cerebellum, hidbrain, middle hind-brain boundary and pharyngeal arches during zebrafish embryogenesis. Knocking down of prmt1 by antisense morpholino oligo results in several morphological defects in embryos in a dose-dependent manner, including malformed tail, shortened and curved body axes, myeloid,yolk shape of change. These data clearly demonstrate a role for PRMT1 in specific developmental processes of zebrafish.

中文摘要.............................................. 1
英文摘要.............................................. 2
第一章 序論........................................... 4
第二章 實驗材料與方法.................................13
第三章 研究結果...................................... 26
第四章 討論.......................................... 33
參考文獻............................................. 37
圖表與圖表說明....................................... 43
附圖................................................. 59

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