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研究生: 陳玉佳
Yu-Chia Chen
論文名稱: 脊索動物AMPA受器次型的基因演化分析
Evolution of the Chordate AMPA Receptor Subunit Genes
指導教授: 周姽嫄 教授
Wei-Yuan Chow
口試委員:
學位類別: 博士
Doctor
系所名稱: 生命科學暨醫學院 - 生命科學系
Department of Life Sciences
論文出版年: 2004
畢業學年度: 92
語文別: 中文
論文頁數: 79
中文關鍵詞: 脊索動物AMPA受器次型基因複製後轉錄作用
外文關鍵詞: chordate, AMPA receptor, gene duplication, post-transcriptional modification
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  • 基因或基因體複製長久以來被認為是使得物種由簡單演化成具有複雜功能的重要機制。過去以RT-PCR的方式從軟骨魚類鯊魚腦選殖出四種AMPA受器次單元,硬骨魚類河豚和斑馬魚各選殖出七種AMPA受器次單元,吳郭魚選殖出八種AMPA受器次單元,四足類青蛙及烏龜腦選殖出四種AMPA受器次單元。本文也以相同的RT-PCR設計,從硬骨魚類演化層次較為原始的長鯙腦選殖出七種AMPA受器次單元;以RT-PCR和genomic PCR從無顎類的盲鰻選殖出兩種AMPA受器次單元;以genomic PCR從頭索動物文昌魚選殖出三種AMPA受器次單元。以哺乳動物AMPA受器次型保留性較高的氨基酸序列作同源性搜尋尾索動物海鞘基因庫,得到一種AMPA受器次單元。將以上脊索動物AMPA受器次型氨基酸序列建立系統發育樹狀圖,由樹狀圖結果推測無脊椎脊索動物至脊椎動物AMPA受器次型基因數目的演變應是源自於單一基因,其後在頭索類及無顎類時發生一至二次基因複製。除此之外,由脊索動物AMPA受器基因結構和後轉錄作用的分析結果得知脊索動物AMPA受器次型譯讀TMD1-TMD4區域的基因結構大致相同,不同的是脊椎動物AMPA受器次型基因結構含有兩個mutually exclusive exons,稱為flop和flip-coding exon,而無脊椎脊索動物卻只有一個flop/flip-coding exon。由脊索動物flop和 flip氨基酸序列建立的系統發育樹狀圖顯示脊椎動物flop與flip-coding exon是經由tandem exon duplication所產生。以Mfold程式預測無脊椎脊索動物海鞘和文昌魚AMPA受器次型基因Q/R site或是R/G site鄰近序列的二級結構,以及將此段序列與哺乳動物AMPA受器基因會進行editing的序列進行比對,結果顯示無脊椎脊索動物AMPA受器次型不會形成editing所需的二級結構,因此無法進行RNA editing。由此推測AMPA受器次型的RNA editing機制只存在脊椎動物中。


    Gene or genome duplications increasing in organismal complexity are important events in evolution. On the basis of RT-PCR surveys, the tetrapod brains and the cartilaginous shark brain expressed four types of AMPAR subunits, and the brains of tilapia, pufferfish and zebrafish expressed eight or seven types of AMPAR subunits. Results of a similar RT-PCR survey showed that the teleost moray eel brain expressed seven kinds of AMPAR subunits, the jawless hagfish brain expressed two types of AMPAR subunits, and the cephalochordate amphioxus had three types of AMPAR subunits. Moreover, using the conserved amino acid sequences of AMPAR subunits as a query to search the urochordate ascidians database, one kind of ascidians AMPAR subunit was obtained. A phylogenetic tree of ascidians, amphioxus, shark, teleost, and tetrapod AMPAR subunits was contracted by Neighbor-joining analysis. This tree suggested that the vertebrate AMPAR subunit genes arose from a common ancestral gene. Subsequently, one or two gene duplication events might occur at either the cephalochordate or the jawless fish lineage. The gene structure of TMD1-TMD4 of chordate AMPAR subunits showed that the vertebrate AMPAR genes included two mutually exclusive exons, named flop and flip. On the other hand, there was only one flop/flip-coding exon in the non-vertebrate chordate AMPAR genes. On the basis of the phylogenetic relationships of flop and flip chordate sequences, the two vertebrate flop and flip-coding exon could result from a primordial exon causing by the tandem exon duplication. The mfold program prediction about the sequences around the Q/R site or the R/G site showed that the non-vertebrate chordate AMPAR transcripts could not form stable secondary structures. This result suggested that the modification of the AMPAR transcripts by Q/R or R/G editing were absent in the non-vertebrate chordate.

    中文摘要………………………………………………I 英文摘要………………………………………………II 前言……………………………………………………1 材料與方法……………………………………………16 結果……………………………………………………26 討論……………………………………………………36 參考文獻………………………………………………45 圖表及附錄……………………………………………56

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    Keenan, Edward L. 本論文從田調語言學和衍生句法學的角度,研究尖石地區賽考利克泰雅語的關係子句。帶有連繫詞ka□的關係子句和不帶有連繫詞ka□的關係子句,證明為兩種不同的關係子句。前者結構上為「外在型關係子句」,語意上為「限制性關係子句」;而後者結構上為「內在型關係子句」,語意上為「非限制性關係子句」。這項區別可以由詞序、接應代詞、時間狀語和專有名詞等證據支持。連繫詞ka□也證明為補語標記,並非關係代名詞或是格位標記。
    基於泰雅語的外在型關係子句,我們主張傳統的「標準分析法」較優於Kayne(1994)所提出的「非對稱分析法」。雖然非對稱分析法可以衍生各種線性次序,但是這種分析不僅會造成過度衍生的現象,更無法解釋賽考利克泰雅語中補語標記的省略。相反地,標準分析法不但沒有這些問題,還能一致性地解釋泰雅語、漢語和英語中外在型關係子句的詞組結構。此外,泰雅語的外在型關係子句呈現出孤島效應上的「論元-附加語不對稱」,以及關係化上的「主賓語不對稱」,這些現象更加支持標準分析法的分析。
    至於泰雅語的內在型關係子句,因為句法上的分佈不相同,所以並不能分析為狀語子句。就孤島效應而論,名前內在型關係子句和名後內在型關係子句在句法上有其不同:前者的中心名詞移位至補語詞組的指示語位置,檢驗時間屬性和關係化屬性;後者的中心名詞則基底衍生在補語詞組的指示語位置。然而,不論名前名後,內在型關係子句都是非限制性關係子句。

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