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研究生:江淑雅
研究生(外文):Shu-Ya Chiang
論文名稱:CTG三聯核酸重複序列擴增對線蟲肌肉功能影響之研究
論文名稱(外文):Study of pathogfenic effects of expanded CTG trinucleotide repeats on the muscle function of C. elegans
指導教授:蕭光明蕭光明引用關係
指導教授(外文):Kuang-Ming Hsaio
學位類別:碩士
校院名稱:中山醫學大學
系所名稱:醫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:44
中文關鍵詞:強直型肌肉萎縮症線蟲CTG三聯核酸重複序列咽喉肌電圖
外文關鍵詞:myotonic dystrophyC. elegansCTG trinucleotide repeatselectropharyngealgram
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(CTG/CAG)n三聯核酸重複序列之擴增突變與多種人類疾病之發生有密切關係。其中,位於染色體19q13.3區域內DMPK基因3’端不轉譯區的此一序列之擴增會造成強直型肌肉萎縮症(myotonic dystrophy)。在此研究中,我們嘗試以線蟲(C. elegans)作為疾病模式動物來研究強直肌肉萎縮症之可能致病機制。首先,我們以PCR-cloning方法將不同長度之CTG三聯核酸重複序列接入pGEM-T質體,再轉至含線蟲肌肉專一性驅動之綠螢光蛋白基因的載體。為模仿人體內真實情形,CTG三聯核酸重複序列係插入綠螢光蛋白基因的3’端不轉譯區。經顯微注射產生之基因轉殖線蟲共有myo3::gfp,myo-3::gfp(CTG)5,及myo-3::gfp(CTG)120三種。其中,myo-3::gfp(CTG)120 line之螢光表現量約只有myo-3::gfp(CTG)5 line之20%以及GFP line之7%,顯示擴增之CTG三聯核酸重複序列會降低其所在基因之表現。 myo-3::gfp(CTG)5 line之C. elegans身體移動速度正常且移動軌跡呈正常規則S型,而myo-3::gfp(CTG)120 line 線蟲之移動則明顯緩慢且不甚規則。此外,我們以線蟲的咽喉肌電圖(EPG)來觀察其肌肉生理發現myo-3::gfp(CTG)120 line之肌肉收縮及舒張明顯異常。這些結果證實擴增之CTG重複序列會造成線蟲肌肉功喪失。

Expansion mutation of CTG repeat length in the 3’-untranslated region of human DMPK gene causes a dominantly inherited muscular disease, called myotonic dystrophy type1 (DM1). In this study we attempt to establish a DM1-disease animal model using transgenic C. elegans. We made transgenic constructs in which (CTG)n was inserted into the 3’-untranslated region of GFP gene driven by muscle-specific promoter of C. elegans. Three lines, including myo-3::gfp, myo-3::gfp(CTG)5, and myo-3::gfp(CTG)120, were generated. As judged from western blotting, the GFP protein level in myo-3::gfp(CTG)120 line is only about 20% of that in myo-3::gfp(CTG)5 line, and is about 7% of that in myo-3::gfp . This result indicates that expanded CTG repeats will greatly decrease the GFP gene expression. In addition, the rate and patterns of body movement as well as electropharyngealgram (EPG) of myo-3::gfp(CTG)120 line is significantly deviated from those observed in myo-3::gfp. myo-3::gfp(CTG)5 line presents normal muscle activity and coordination. These data demonstrate that expanded CTG repeats have pathogenic effects on C. elegans

目次
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中文摘要………………………………………………… 2
英文摘要………………………………………………… 3
序 論………………………………………………… 4
材料與方法……………………………………………… 11
結 果………………………………………………… 23
討 論………………………………………………… 27
圖 表………………………………………………… 31
參考文獻………………………………………………… 39

參考文獻
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