- 學位論文
多品系基因改造玉米檢測方法之開發
Development of detection methods for Multiline Genetically Modified Maize
摘要
目前全球基因改造生物(Genetically Modified Organisms, GMO)發展，依然快速增長中，根據Agbios 資料庫顯示2008年全球核准上市的基因改造玉米共有44種，其中約有四分之一屬於混合型(stacked)品系的玉米，這些新品系最大的爭議在於是否須要重新評估其安全性。我國目前核准基因改造玉米品系則為單一基因改造玉米品系14種，尚未核定任何混合品系，為釐清我國目前市場可見之販售玉米與核定品系間之遵循性，以做為對基改食品管理機制的基礎，故進行此調查研究，本實驗共分成基改玉米鑑定系統與基改玉米定量系統兩部份。 基改玉米鑑定系統方面，由全臺灣共取得107個取樣點取的顆粒完整之散裝玉米樣品，由樣品中選取共321個試驗樣品，進行基改玉米篩選。35S 啟動子、nos 終止子及玉米內源性基因zein進行初篩，結果共獲得17個基因改造玉米的陽性樣品，再以Onishi等學者於2005年所發表之基改玉米鑑定系統進行品系檢定，在17個陽性樣品中，篩得9個含有基因改造成分，包含NK603品系、NK603ＸMON810混合品系、NK603ＸMON863混合品系、NK603、MON863與M0N810混合品系、MON863品系等；基改玉米定量系統方面則以以Yang等學者於2007年與Kuribara等學者於2002所發表之研究報告為基礎，配合所能取得之基改玉米標準品，建立SYBR Green I 定量系統之建立，認證標準樣品包含品項混雜比例從0~5.0%不等，經過驗證後此定量系統的偏估值變動的分布主要在± 41%以內，而變異係數變動範圍主要在33%以內，顯示此定量系統可得到可靠的定量結果。
並列摘要
Currently, there is a rapid development on Genetically Modified Organisms(GMO) globally. Fourty-four kinds of GM maize have been approved to launch to the market in 2008 according to Agbios database. Appropriraely one quarter of those 44 kinds are Stacked maize. There are many disputes on the necessity for re-evaluating the safety of these new products currently. Taiwan government has not approve amy imported Stacked maize. In order to provide a basic GM-food managing mechanism, clarifying the relationship between the maize in the market and approved GM maize lines is im-portant. This study consists of two parts, namely, GMO screening PCR system and GM maize quantitative detection. In the GMO screening PCR system, the bulk maize samples intact which 107 sam-pling in Taiwan, and 321 test samples were chosen from the sample to select GM maize. Using 35S promoter, NOS terminator and maize endogenous gene zein to proceed the first stage of selection and we get 17 GMO positive GM maize. Then, based on the “GMO screening PCR system” published by scholars including Onishi in 2005, GMO positive maize out of 17 GMO positive samples were selected, including, NK603lines、NK603ＸMON863 stacked lines、NK603ＸMON810 stacked lines、NK603、MON863 and M0N810 stacked lines、MON863 lines. The GM maize quantitative detection is based on two published reports. Using the certified reference materials include certified reference materials ranging varies from 0~5.0% to develop real-time PCR systems, which is based on SYBR Green I technologies for specific guantitative. In this experi-ment, the bias range of quantification system has been approved within ±33%, the coef-ficients of variation distribute within 41%. These results have proved that this GMO quantification system developed in this study is reliable in practice.