臺灣博碩士論文加值系統

背景與目的 選擇性剪裁是真核細胞生物由pre-mRNA產生mRNA的重要步驟.真核細胞在基因的表現上由於選擇性剪裁的緣故，能產生更多種類的蛋白質，也是生物於不同組織，不同生長時期控制gene表現的重要調控機制。選擇性剪裁產生的mRNA可以轉譯出蛋白質以維持生物細胞生理功能，其異常亦可造成疾病 許多公開可用的分子生物學資料庫隱含極有用的資訊。 Xie H.等曾以EST比對unigene的選擇性剪裁資料庫尋找與腫瘤，組織，有關的基因與選擇性剪裁樣式，其文章中只簡要探討直腸癌的選擇性剪裁。精神分裂症與生長發育目前尚未有利用計算科學篩選選擇性剪裁的研究。本文即是以EST資料庫計算出的選擇性剪裁資訊¸探討其與疾病與生長階段的關聯。
方法
資料來源: EST資料與組織來源library資料，疾病資料庫OMIM下載自NCBI網站。細胞生理流程pathway資料庫KEGG，蛋白質資料庫下載自各別網站。蛋白質資料猜拆解於genbank檔案。各檔案以perl或C++程式拆解，輸入IBM DB2及MS-access。
資料處理: 1．建立選擇性剪裁資料庫(AVATAR):本實驗室經由自行發展的多層單標籤演算法將EST比對到全基因體的序列上，建立intron，exon，以及五種選擇性剪裁樣式的資料庫。 2．找出疾病與生長有關的選擇性剪裁位置:將組織來源資料庫依特定腫瘤(包括前十大癌症)，精神分裂症，及生長階段予以編碼。再將每個選擇性剪裁位置上比對到的EST個數，依isoform種類， 是否為某種疾病計算其發生頻率，以Fisher exact test檢驗其差異顯著性，探討此位置是否有疾病或生長階段有關。 3．將有關的選擇性剪裁位置，依蛋白質與基因位置找出其相對映的蛋白質與基因，並建立一依基因為串聯索引，包含疾病， 細胞生理流程，選擇性剪裁的綜合資料庫。
結果 依各疾病種類，我們找到了各選擇性剪裁樣式有關的選擇性剪裁位置。例如在精神分裂証方面，5’選擇性剪裁及exon skip樣式各有三處與疾病有關，mutual exclusion有四處。肝癌方面，exon skip有63處，intron retain有231處選擇性剪裁位置與疾病有關。與生長發育時期有關的選擇性剪裁位置共有163處
討論 找到的選擇性剪裁位置需進一步於實驗室中驗證。結合現有疾病，蛋白質，細胞生理流程的資料，將可篩選出較有意義的位置，並進一步探討其與疾病發生的可能機轉，將有助於臨床診斷與治療。

Introduction:
Alternative splicing is important because more proteins can be made from the relative few genes and. it is also an important mechanism regulating genetic expression. In this thesis, we will try to find out alternative splicing isoforms specific to several cancers and schizophenia and different development stage. The integrated information about diseases, pathways, proteins, and alternative splicing will be present.
Methods:
Alternative splicing database were constructed by aligning EST to whole genomic sequence using multi-layer unique marker methods. Each splicing site will be queried for their EST expression frequency in specific disease or development stage by the specific splicing isoforms. Significance were tested with Fish’s exact test. Using the position in the contig, splicing site was link to their correlated pretein, gene. By linking the same gene, disease information in OMIM and pathway information in KEGG were integrated.
Results:
1263 disease specific splicing sites were identified. 63 exon skipping sites and 231 intron retain sites were correlated with hepatoma. 17 sites were correlated with schizophrenia. 163 splicing site were specific to fetal and infantile development stage.
Discussion:
Further laboratory investigation will be needed for confirming the real significant splicing site clinically.

1. 封面
2. 口試委員簽名頁
3. 中文摘要
4. 英文摘要
5. 目錄
6. 表目錄
7. 論文本文
8. 參考文獻
9. 誌謝
10. 簡歷

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