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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 徐源泰 | |
| dc.contributor.author | Ya-Peng Hou | en |
| dc.contributor.author | 侯俹芃 | zh_TW |
| dc.date.accessioned | 2021-06-13T06:03:07Z | - |
| dc.date.available | 2006-07-28 | |
| dc.date.copyright | 2006-07-28 | |
| dc.date.issued | 2006 | |
| dc.date.submitted | 2006-06-20 | |
| dc.identifier.citation | 王慶裕，2000，固殺草 (glufosinate) 除草劑之作用及抗性機制，
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/34324 | - |
| dc.description.abstract | 近年來世界各國逐漸重視基因改造食品的應用和安全性，並制定相關標準與法規予以規範與管理，我國基因改造食品於西元2003年起實行強制標示。本研究以35S-F/35-R和nos-F/nos-R兩組引子進行複式聚合酶鏈鎖反應，可正確檢測出一種基因改造大豆及四種基因改造玉米。針對基因改造玉米Bt11和Bt176之轉殖基因3’端與玉米染色體DNA相接處 (integration site) 設計具品系專一性 (event-specific) 之引子Bt11-F/Bt11-R、Bt176-F/Bt176-R；針對基因改造玉米MON810和GA21之基因改造5’端與玉米染色體DNA相接處設計具專一性之引子MON810-F/MON810-R和GA21-F/GA21-R。以即時聚合酶鏈鎖反應TaqMan進行定量，四種基因改造玉米品系Bt11、Bt176、MON810、GA21之平均內標值分別約為0.74、0.62、0.26、1.41；定量檢測系統之正確性檢驗，CV範圍介於3.69 ~ 35.81%間，偏移值介於0.08 ~ 0.24%間。針對基因改造大豆RRS之轉殖基因3’端與大豆染色體DNA相接處設計具有專一性之引子RRS-F/RRS-R；以即時聚合酶鏈鎖反應SYBR Green I dye進行定量，基因改造大豆RRS之平均內標值約為1.15。大量樣品取樣方法，設計出三種類型20種取樣方法：類型Ⅰ比較不同樣本大小；類型Ⅱ比較階段式取樣之差異；類型Ⅲ比較複合樣品間之差異，類型Ⅲ取樣方法17複合樣品以三點取樣，對於原始族群也較具有代表性，檢驗上也較省時低成本，此方法可作為日後大量檢體取樣方法之參考。 | zh_TW |
| dc.description.abstract | The detection of GM-food has its necessity due to food safety regulation, and the labeling of GM-food has been practiced since 2003 in Taiwan. The use of multiplex PCR containing 35S-F/35S-R and nos-F/nos-R is found applicable to detect four lines of genetically modified maize: Bt11, Bt176, MON810, GA21 maize, and one line of genetically modified soybean: Roundup Ready soybean. We further designed the event-specific primers to target the 3’ integration sites of Bt11 and Bt176 and the 5’ integration sites of MON810 and GA21. The quantification of Bt11, Bt176, MON810 and GA21 were carried out by using Lightcycler Instrument with TaqMan Kit. The coefficient values of Bt11, Bt176, MON810 and GA21 were 0.74, 0.62, 0.26 and 1.41. The accuracy of real-time Q-PCR detection methods, expressed as coefficient of variation for Bt11, Bt176, MON810 and GA21 varied from 3.69% to 35.81% and bias range from 0.08% to 0.24%. We designed the event-specific primers to target the 3’ integration sites of RRS. We developed the event-specific real-time detection and quantification of RRS using Lightcycler Instrument with SYBR Green I dye. The coefficient value of RRS was 1.15. In the development of bulk sampling methods, we designed twenty sampling methods in three types. In type Ⅰ, different sample sizes were compared; in type Ⅱ, we compare the process of multi-stage sampling; in type III, the mixed sampling methods were studied. The results showed that the sampling method 17 in type Ⅲ could decrease variations between individuals and save more time and cost. This method is found potential to be applied in bulk sampling process in future. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T06:03:07Z (GMT). No. of bitstreams: 1 ntu-95-R93628201-1.pdf: 3680776 bytes, checksum: 57bae2a74176a40d72abf19e8119a468 (MD5) Previous issue date: 2006 | en |
| dc.description.tableofcontents | 頁次
目錄 Ⅰ 圖次 Ⅴ 表次 Ⅸ 摘要 ⅩⅠ Abstract ⅩⅡ 壹、前言 1 貳、前人研究 3 一、基因改造生物之概說 3 (一)什麼是基因改造生物 3 (二)基因改造生物的運用與功能 3 (三)基因改造作物之現況 4 (四)基因改造食品安全性 10 (五)各國基因改造產品規範 11 二、基因改造樣品之檢驗 12 (一)基因改造樣品之定性檢測 12 (二)基因改造樣品之定量檢測 17 A. 定量競爭聚合酶鏈鎖反應 17 B. 即時定量聚合酶鏈鎖反應 18 (三)大量檢體之取樣 18 (四)統計分析軟體SAS 19 三、基因改造大豆之介紹 25 (一)大豆簡介 25 (二)抗嘉磷塞黃豆 (Roundup ReadyTM soybean, RRS) 介紹 26 A. Roundup殺草劑之作用機制 26 B. 抗嘉磷賽除草劑大豆之特性 27 C. 大豆內源性基因 27 四、基因改造玉米之介紹 29 (一)玉米簡介 29 (二)基因改造玉米介紹 29 A. 生物性殺蟲劑介紹 30 B. 固殺草劑(glufosinate ammonium)之介紹 33 C. 玉米內源性基因 33 叁、材料與方法 42 一、研究材料 42 二、藥品 42 三、儀器設備 43 四、實驗方法 44 (一)基因改造玉米 44 A. DNA的製備與純化 44 B.標準參考質體的建立 45 C.定性檢測定 46 D. 即時定量聚合酶鏈鎖反應 (Lightcycler定量分析) 47 (二)基因改造大豆 48 A. 基因改造大豆取樣方法 48 B. DNA的製備與純化 52 C.標準參考質體的建立 52 D.定性檢測定 53 E. 即時定量聚合酶鏈鎖反應 (Lightcycler定量分析) 54 F. 統計分析 54 肆、結果與討論 59 一、基因改造玉米 59 (一)基因改造玉米之定性分析 59 A.以Ivr-F/Ivr-R引子進行玉米之定性分析 59 B.以Bt11-F/Bt11-R引子進行基因改造玉米Bt11之定性分析 59 C.以Bt176-F/Bt176-R引子進行基因改造玉米Bt176之定性分析 60 D.以MON810-F/MON810-F引子進行基因改造玉米MON810之 定性分析 60 E.以GA21-F/GA21-R引子進行基因改造玉米GA21之定性分析 60 (二)複式聚合酶鏈鎖反應 61 (三) 即時聚合酶鏈鎖反應檢測系統的建立 70 A.標準參考質體的構築 71 B.標準曲線與檢測範圍 72 C.內標比(coefficient value, Cv) 90 D.檢測系統的正確性(accuracy) 91 二、基因改造大豆 94 (一)基因改造大豆RRS定行分析 94 A.以LECTIN-F/LECTIN-R引子對進行大豆之定性分析 94 B.以RRS引子對進行基因改造大豆RRS之定性分析 94 (二)複式聚合酶鏈鎖反應 98 (三) 即時聚合酶鏈鎖反應檢測系統的建立 98 A.即時聚合酶鏈鎖反應檢測之定性分析 98 B.標準參考質體的構築與檢測範圍 98 C.內標比 99 D.檢測系統的正確性 100 (四)基因改造大豆取樣之定量結果 107 A.類型Ⅰ取樣方法1-8定量結果之比較 107 B.類型Ⅱ取樣方法9-12定量結果之比較 109 C.類型Ⅰ和類型Ⅱ取樣方法1-12定量結果之比較 109 D.類型Ⅲ取樣方法13-20定量結果之比較 110 E.類型ⅠⅡⅢ之綜合比較 110 伍、結論 117 陸、參考文獻 119 | |
| dc.language.iso | zh-TW | |
| dc.title | 基因改造大豆及四種品系玉米檢測方法之研究 | zh_TW |
| dc.title | Study on the detection methods of genetically modified soybean and maize in four lines | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 94-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 陳昭瑩,鄭石通,許輔 | |
| dc.subject.keyword | 基因改造大豆,基因改造玉米,即時定量聚合酶,鏈鎖反應, | zh_TW |
| dc.subject.keyword | GMO,RRS,Bt11,Bt176,MON810,GA21,real-time PCR, | en |
| dc.relation.page | 131 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2006-06-21 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 園藝學研究所 | zh_TW |
| 顯示於系所單位： | 園藝暨景觀學系 | |
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