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研究生:張肇源
研究生(外文):Chao-Yuan Chang
論文名稱:利用狐尾草嵌紋病毒擴展竹嵌紋病毒衛星核酸寄主範圍之可行性研究
論文名稱(外文):Study on the Potentiality of Using Foxtail Mosaic Virus to Extend the Host Range of Satellite RNA of Bamboo Mosaic Potexvirus
指導教授:徐堯煇
指導教授(外文):Yau-Heiu Hsu
學位類別:碩士
校院名稱:國立中興大學
系所名稱:農業生物科技學研究所
學門:農業科學學門
學類:農業技術學類
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
中文關鍵詞:狐尾草嵌紋病毒竹嵌紋病毒衛星核酸
外文關鍵詞:Foxtail Mosaic VirusSatellite RNA of Bamboo Mosaic Potexvirus
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竹嵌紋病毒 (Bamboo mosaic potexvirus, BaMV) 屬於馬鈴薯X病毒群 (Potexvirus group),有些BaMV分離株攜帶有衛星核酸 (satellite RNA, satBaMV),SatBaMV RNA為單股、正極性,不包括poly(A),全長為836 個核苷酸,含有一個轉譯區,可表現一個非結構性的20 kDa蛋白質 (P20),此轉譯區非複製所必須,可為外源基因取代,已發展成載體系統。BaMV可感染之植物僅包括竹、水稻、大麥及菸草 (Nicotiana benthamiana),其寄主範圍過於狹窄為衛星核酸載體其應用的限制因素。同屬於馬鈴薯病毒群的狐尾草嵌紋病毒 (Foxtail mosaic virus, FoMV),其寄主範圍包括數十種單子葉植物及雙子葉植物。實驗結果已知將FoMV與satBaMV RNA共同接種菸草的原生質體,FoMV亦可以幫助satBaMV RNA進行複製,而其P20之表現量約為以BaMV協助時的五分之一。為利用狐尾草嵌紋病毒寄主範圍較竹嵌紋病毒寄主範圍廣泛的特性,來擴展竹嵌紋病毒衛星核酸之寄主範圍,本實驗首先構築狐尾草嵌紋病毒具感染力之cDNA選殖株,並測試其寄主範圍,發現此選殖株主要感染禾本科、茄科植物,但對於十字花科植物不具感染力。同時,利用此選殖株接種satBaMV RNA全長轉殖植物時,可以測得P20表現大量增加,表示此選殖株能夠幫助SatBaMV RNA進行複製。此外,本實驗亦利用FoMV及BaMV的cDNA株構築雜合病毒﹙chimeric viruses﹚,先將可轉譯出複製酵素之第一個轉譯區﹙ORF 1﹚進行置換,再與satBaMV RNA共同接種各種植物,以進行感染力的測試。結果顯示以狐尾草嵌紋病毒的cDNA株作為骨幹,置換竹嵌紋病毒之第一個轉譯區的重組株無法進行有效複製,但以竹嵌紋病毒的cDNA株作為骨幹,置換狐尾草嵌紋病毒之第一個轉譯區的重組株則可以進行複製。未來研究可繼續探討此具感染力之雜合病毒株是否能擴展竹嵌紋病毒衛星核酸載體應用於BaMV寄主外之植物。

壹、中文摘要 ----------------------------------------------- 2
貳、英文摘要 ----------------------------------------------- 3
參、前言 --------------------------------------------------- 4
肆、材料與方法 --------------------------------------------- 8
1. 狐尾草嵌紋病毒具感染力cDNA選殖株之構築 ------------------ 8
2. 狐尾草嵌紋病毒多株抗體之製備 ---------------------------- 12
3. 狐尾草嵌紋病毒具感染力cDNA選殖株之核苷酸定序 ------------- 12
4. 狐尾草嵌紋病毒選殖株寄主範圍之測試 ---------------------- 14
5. 狐尾草嵌紋病毒選殖株與竹嵌紋病毒衛星核酸共同接種之測試 -- 16
6. FoMV與BaMV第一個轉譯區互換之雜合病毒之構築 --------------- 19
7. 原生質體接種測試 ---------------------------------------- 20
伍、結果 --------------------------------------------------- 24
1. 篩選具有感染力之狐尾草嵌紋病毒cDNA選殖株 ---------------- 24
2. 狐尾草嵌紋病毒具感染力cDNA選殖株之核苷酸序列分析 --------- 24
3. 狐尾草嵌紋病毒選殖株之寄主範圍 -------------------------- 25
4. 狐尾草嵌紋病毒選殖株與竹嵌紋病毒衛星核酸共同接種植株之測試分
析 ------------------------------------------------------- 27
5. FoMV與BaMV第一個轉譯區互換之雜合病毒之感染力 ------------ 28
陸、討論 --------------------------------------------------- 30
柒、參考文獻 ----------------------------------------------- 35
捌、圖表、附錄 --------------------------------------------- 41

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