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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 胡凱康(Kae-Kang Hwu) | |
dc.contributor.author | Chung-Ho Lei | en |
dc.contributor.author | 雷中和 | zh_TW |
dc.date.accessioned | 2021-06-13T00:05:03Z | - |
dc.date.available | 2007-08-02 | |
dc.date.copyright | 2007-08-02 | |
dc.date.issued | 2007 | |
dc.date.submitted | 2007-07-27 | |
dc.identifier.citation | 行政院衛生署食品藥物檢驗局-基因改造食品資訊網:GM食品標示, 2007, http://food.doh.gov.tw/chinese/info/gmo4.htm
食品資訊網–業務資訊查詢, 2007, http://food.doh.gov.tw/chinese/info/gmo4.htm 侯俹芃,2006,基因改造大豆及四種品系玉米檢測方法之研究,國立台灣大學園藝系碩士論文。 Afonina, I., M. Zivarts, I. Kutyavin, E. Lukhtanov, H. Gamper, and R.B. Meyer. 1997. Efficient priming of PCR with short oligonucleotides conjugated to a minor groove binder. Nucleic Acids Res 25:2657-60. Agbios , GM database, 2007, http://www.agbios.com/dbase.php An, Y., J. Ji, W. Wu, A. Lv, R. Huang, and Y. Wei. 2005. A rapid and efficient method for multiple-site mutagenesis with a modified overlap extension PCR. Appl Microbiol Biotechnol 68:774-8. Bennett, M. 1995. Nuclear and chromosomal DNA amounts in Zea mays spp. mays. http://www.maizegdb.org/ancillary/zmdnaamt.html Bennett, M., and J. Leitch. 2005. Plant DNA C-values Database http://www.kew.org/cval/database1.html Block, A., and G. Schwarz. 2003. Validation of different genomic and cloned DNA calibration standards for construct-specific quantification of LibertyLink in rapeseed by real-time PCR. European Food Research and Technology 216:421-427. Bruderer, S., and E. Leitner K. 2003. Genetically modified (GM) crops : molecular and regulatroy details http://www.bats.ch/gmo-watch/GVO-report140703.pdf Hernández, M., T. Esteve, S. Prat, and M. Pla. 2004. Development of real-time PCR systems on SYBRw Green I, Amplifluore and TaqManw technologies for specific quantitative detection of the transgenic maize event GA21. Journal of Cereal Science 39:99-107. Holst-Jensen, A., S.B. Ronning, A. Lovseth, and K.G. Berdal. 2003. PCR technology for screening and quantification of genetically modified organisms (GMOs). Anal Bioanal Chem 375:985-93. Huang, C., T. Shih, and T. Pan. 2004. Development and application of a nested polymerase chain reaction method for the detection of genetically modified Soybean in Chinese traditional fermented soy food-sufu. Journal of Food and Drug Analysis 12:266-272 James, C,. 2006. Global Status of Commercialized Biotech/GM Crops: 2006 http://www.isaaa.org/Resources/Publications/briefs/35/pptslides/Brief35slides.pdf James, D., A.M. Schmidt, E. Wall, M. Green, and S. Masri. 2003. Reliable detection and identification of genetically modified maize, soybean, and canola by multiplex PCR analysis. J Agric Food Chem 51:5829-34. Kuribara, H., Y. Shindo, T. Matsuoka, K. Takubo, S. Futo, N. Aoki, T. Hirao, H. Akiyama, Y. Goda, M. Toyoda, and A. Hino. 2002. Novel reference molecules for quantitation of genetically modified maize and soybean. J AOAC Int 85:1077-89. Kutyavin, I.V., E.A. Lukhtanov, H.B. Gamper, and R.B. Meyer. 1997. Oligonucleotides with conjugated dihydropyrroloindole tripeptides: base composition and backbone effects on hybridization. Nucleic Acids Res 25:3718-23. Lee, S., S. Kang, Y. Park, D. Min, and Y. Kim. 2006. Quantitative analysis of two genetically modified maize lines by real-time PCR. J. Microbiol. Biotechnol. 16:205-211. Matsuoka, T., H. Kuribara, K. Takubo, H. Akiyama, H. Miura, Y. Goda, Y. Kusakabe, K. Isshiki, M. Toyoda, and A. Hino. 2002. Detection of recombinant DNA segments introduced to genetically modified maize (Zea mays). J Agric Food Chem 50:2100-9. Onishi, M., T. Matsuoka, T. Kodama, K. Kashiwaba, S. Futo, H. Akiyama, T. Maitani, S. Furui, T. Oguchi, and A. Hino. 2005. Development of a multiplex polymerase chain reaction method for simultaneous detection of eight events of genetically modified maize. J Agric Food Chem 53:9713-21. Querci, M., M. Jermini, and G.V.d. Eede. 2004. The analysis of food samples for the presence of genetically modified organisms : user manual. http://gmotraining.jrc.it/manual.htm, Trapmann, S. 2006. Use of certified reference materials for the quantification of GMO in food and feed. European Reference Material. http://www.irmm.jrc.be/html/reference_materials_catalogue/user_support/erm_application_notes/application_note_4/application_note_4_english.pdf Weighardt, F., C. Barbati, C. Paoletti, M. Querci, S. Kay, M. De Beuckeleer, and G. Van den Eede. 2004. Real-time polymerase chain reaction-based approach for quantification of the pat gene in the T25 Zea mays event. J AOAC Int 87:1342-55. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/28317 | - |
dc.description.abstract | 本研究目的在於建立能檢定基因改造玉米混雜率的定量系統,以聚合酵素連鎖反應為基礎技術的即時聚合酵素反應,針對Bt11、176、GA21、NK603、MON810、MON863、TC1507等七個基因改造玉米品項設計具有專一性的定量系統。首先以定序取得七個基因改造玉米品項的部份轉殖基因序列,進而針對其中具有專一性的片段來作為設計引子對及探針,並將所挑選的專一性片段加上玉米內生基因片段zSSIIb、轉殖基因通用啟動子P35S與終結子Tnos接合轉殖進質體當作定量用途的標準物質,最後實際以認證標準物質 (certified reference material, CRM) 驗證本研究中建立的定量系統定量的精準度。認證標準樣品包含各品項混雜比例從0~10.0%不等,經過驗證後此定量系統的偏估值變動的分布主要在 %以內而變異係數變動範圍主要在20%以內,顯示此定量系統可得到可靠的定量結果。 | zh_TW |
dc.description.abstract | The purpose of this study is to establish a quantification system based on real-time PCR for 7 most common genetically modified events of maize. Targeted events include Bt11, 176, GA21, NK603, MON810, MON863 and TC1507. Construct specific primers and probes were designed according to the partial sequences of inserts verified by direct sequencing of PCR amplicons of primers found in the literatures. Specific amplicons of the above mentioned events and maize endogenous DNA sequence zSSIIb, cauliflower mosaic virus 35S promoter, nopaline synthase terminator were appended and inserted into PGEM-T vector. A PstI digested linear form of transformed vector was quantified and used to establish the standard curves of each event. Certified reference materials ranging from 0 to 10% were used to validate the accuracy and precision of this quantification system. In most of the cases, the bias ranged from -30% to 30% and coefficients of variation were less then 20%. These results have proved that the GMO quantification system developed in this study is reliable in practice. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T00:05:03Z (GMT). No. of bitstreams: 1 ntu-96-R93621118-1.pdf: 1372950 bytes, checksum: 8b298f5a27cad8fbb271c826c431870e (MD5) Previous issue date: 2007 | en |
dc.description.tableofcontents | 內容目錄 i
圖片目錄 iii 表格目錄 iv 摘要 v Abstract vi 第一章 前言 1 第二章 前人研究 4 第三章 材料與方法 8 壹、 研究材料 8 貳、 DNA萃取 8 參、 轉殖基因序列之確認 9 一、 引子 (primer) 9 二、 聚合酵素連鎖反應 10 三、 定序以及設計引子 (sequence and primer design) 11 肆、 質體標準樣品之建立與生產 11 一、 質體標準樣品之建立 11 1. 引子及探針的設計 (primer and probe design) 11 2. 引子專一性測試 12 3. 重疊延伸聚合酵素連鎖反應 (overlap extension PCR) 12 二、 質體標準樣品的生產 14 1. 轉殖 (clone) 14 2. 質體DNA的純化 15 3. 限制酶切 15 4. 配製標準樣品 15 伍、 定量系統之驗證 16 一、 質體標準樣品對質體標準樣品 16 二、 質體標準樣品對認證標準物質 16 第四章 結果 18 壹、 轉殖基因序列之確認 18 貳、 質體標準樣品之建立 19 一、 引子與探針的設計 19 二、 引子專一性測試 20 三、 重疊延伸聚合酵素連鎖反應 21 參、 質體標準樣品對質體標準樣品 22 肆、 質體標準樣品對認證標準樣品 22 第五章 討論 26 壹、 定量系統的專一性 26 貳、 質體標準樣品之建立 26 參、 定量系統的精密度 27 肆、 轉換係數的估算 28 伍、 認證標準物質對定量系統的驗證 31 陸、 定量系統的實際應用 33 第六章 結論 36 第七章 參考文獻 38 第八章 附錄 40 附錄一:質體標準物質序列與引子探針黏合位置 40 附錄二:定量系統的標準曲線與精密度參數列表 42 附錄三:定量數據與實際數值的回歸線 47 | |
dc.language.iso | zh-TW | |
dc.title | 研發定量分析系統以檢定基因改造玉米 | zh_TW |
dc.title | Development of quantitative system for detecting GM maize | en |
dc.type | Thesis | |
dc.date.schoolyear | 95-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 潘子明(Tzu-Ming Pan),劉麗飛(Li-Fei Liu),林順福(Shun-Fu Lin) | |
dc.subject.keyword | 基因改造玉米,即時聚合酵素連鎖反應,定量系統, | zh_TW |
dc.subject.keyword | genetically modified maize,real-time PCR,quantitative system, | en |
dc.relation.page | 39 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2007-07-30 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 農藝學研究所 | zh_TW |
顯示於系所單位: | 農藝學系 |
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