本研究室以莫三比克口孵吳郭魚 (Oreochromis mossambicus) 為神經發育研究的模式生物，並探討神經發育相關基因對於神經網絡型塑之影響。本研究室所建構之魚腦發育初期表現序列標記 (Expressed Sequence Tags) 資料庫中，共有1092條序列 (transcript)。本論文僅針對其中一條具有功能性的神經發育相關基因進行研究。Chimaerin 1 (CHN1) 基因在神經系統發育初期扮演重要角色，具影響軸突 (axon) 和樹突 (dendrites) 的生長。本實驗利用生物資訊分析的方法，針對CHN1基因進行開放閱讀區 (Open Reading Frame, ORF) 分析，結果得知CHN1基因ORF之全長為1005 bp 的DNA序列。進一步，建構原核系統 (E. coli) 以進行表現CHN1 protein，並探討CHN1蛋白質對於小鼠神經母瘤細胞 (Neuro-2a, N2a) 在生長上的影響。結果顯示CHN1蛋白質具影響Neuro2a細胞增生的作用。另一方面，應用轉染 (transfection) 技術，將帶有CHN1基因的pEGFP質體送入N2a細胞中，以探討CHN1蛋白質在細胞中的分佈位置，研究結果顯示CHN1蛋白質會分佈在N2a的細胞質中。

Oreochromis mossambicus is as a model organisms for researching molecular neural development biology in our research laboratory. A unique gene chimaerin (CHN1) out of the database of 1092 ESTs which is development tilapia brain in our lab. This thesis just focuses the analysis and research on one of those ESTs that is with the function of CHN1, a neural development-related gene. CHN1 gene play the leading role in the early development of nervous system and influence the growth of axons and dendrites. Bioinformatics were searched for NCBI GenBank: EE724101.1 which is a gene with 1005 bp of open reading frame (ORF), and deduced as 334 amino acid of protein sequence. These results show that CHN1 protein is mainly distributed in the cytoplasma of Neuro-2a cell. The CHN1 reconbinant protein was expressed in E.coli, a prokaryotic system , and purified by Ni-NTA affinity chromatograph. The cellular localization of CHN1 protein was investigated by Neuro-2a cell. The recombinant CHN1 protein has an effect to influence the proliferation of Neuro2a cells.

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致謝 ····································································································· ii
中文摘要 ····························································································· iii
英文摘要 ····························································································· iv
目錄 ····································································································· v
圖次 ····································································································· vi
前言 ····································································································· 1
材料與方法 ························································································· 3
結果 ····································································································· 30
討論 ····································································································· 34
參考文獻 ····························································································· 36

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