Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/56679
標題: 聚合脢鏈反應技術於基因改造玉米之半定量及品系鑑定方法之開發與檢測
Application of Polymerase Chain Reaction on Semi-Quantitative Analysis and Identification of Genetically Modified Corns
作者: 方繼
關鍵字: 應用研究
Genetically modified food
食品科技, 生物技術
基因改造食品
多套式PCR
半定量PCR
限制切割酵素
multiplex PCR
semi-quantitative PCR
restriction enzyme
摘要: 基因改造生物(Genetically Modified Organisms, GMO)於近年來備受關注,其相關研究亦日益增多,目前已有不少基因改造作物於市場上流通。我國進口之玉米中已逾30%為基因改造玉米(http://www.coa.gov.tw),截至九十二年七月底,共有十一種基因改造品系於國內衛生署進行食品安全申請,包括有十種轉殖玉米品系。為有效管理GMO,行政院衛生署目前已公告「基因改造食品安全性評估方法」及「基因改造食品標示辦法」,為我國基因改造食品之管理準則(http://food.doh.gov.tw)。目前基因改造食品之種類日益繁多,雖有許多方法均可檢測基改食品之存在,然國際間仍缺乏一套驗證食品中是否含有基因改造生物檢測方法之制度。本研究室於行政院衛生署之補助下,於九十二年一月起進行多套式聚合酶鏈鎖反應(Multiplex PCR)鑑定方法之開發及建立比對實驗室,評估該技術應用於檢驗基因改造大豆及市售產品之可行性。本年度則擬針對基因改造玉米之半定量分析及品系鑑定進行研究,實驗過程為針對具有cryIA(b)抗蟲基因特性之基因改造玉米其轉殖基因設計不同引子,分別為35SP-F/R之143 bp及文獻已發表cryIA-4-5’/4-3’之152 bp(Matsuoka, et al., 2002),並利用INV-A/B之122 bp作為品種特性基因之鑑定(Vollenhofer, et al., 1999)。此外,為瞭解市售玉米產品中之GMO比例,本實驗亦將市售產品之PCR產物量與不同GMO比例之DNA標準曲線作比對並進行半定量分析,以推斷市售產品中之GMO比例。 本申請案擬以PCR技術(定性及半定量方式)建立基因改造玉米檢驗流程及品系鑑定方法開發,除蒐集國內外已發表檢測基因改造食品之定量方法與本研究室所開發之半定量PCR進行比對及再驗證外,並輔以限制切割酵素反應進行多種基因改造玉米品系之區分及各品系玉米於市售食品之分佈統計。擬利用本研究室開發PCR檢測技術所得之半定量檢測結果,與real-time PCR定量檢測方法所得結果作比較,藉以評估所開發半定量檢測方法之精確性。期藉由此半定量及基因改造玉米品系區分檢驗方法之建立,希能對基因改造食品之檢驗、定量與標示有所助益。
Genetically modified organism has received much attention in resent years and researchers have put their time and efforts in this promising area. In Taiwan, more than 30% of imported corns are genetically modified soybeans. On July 2003, eleven kinds of GMO, including 10 species of genetically modified corns, have applied for safety evaluation by Department of Health (DOH). DOH has promulgated ?HHH?HH?H?HGuidance of Safety Assessment for Genetically Modified Foods?HHH?HHH and ?HHH?HH?H?HLabeling of GM Foods?HHH?HHH that forms the foundation of administration of GM foods in Taiwan.Various methods for detection of GMO have been developed by different researchers and organizations around the world; however, very few validation or collaboratory studies of these methods are available yet. Since the promulgation of these acts mentioned above, effective and correct methods to detect GMO are necessary for DOH in Taiwan. The establishment of collaboratory laboratories and validation of the detection methods for GMO will enhance the standard operation procedures among these institutes. The Applied microbiology and biotechnology laboratory (National Chung Hsing University) has started the development of multiplex PCR technique to detect the GM soybean in feed and food by the financial support of DOH (Taiwan) since 1992. The objective of this study was to develop a semi-quantitative method to detect GM corns based on PCR principle. CryIA(b) (143 bp) and cryIA-4-5?HHH?HHH/4-3 (152 bp). INV-A/B will be used as an identification gene for maize specie. This proposal intends to apply the qualitative and semi-quantitative PCR techniques for establishment the collaboratory laboratory among several institutes in Taiwan. Detection methods for GMO will be collected around the world and compared to the methods that have been developed by our laboratory. In addition, restriction enzyme digestion analysis for distinguishing various strains among GM corns will be conducted. The semi-quantitative PCR that developed by our laboratory and real-time PCR will be compared to validate the accuracy of the semi-quantitative PCR.
URI: http://hdl.handle.net/11455/56679
其他識別: DOH93-TD-F-113-012
文章連結: http://grbsearch.stpi.narl.org.tw/GRB/result.jsp?id=909903&plan_no=DOH93-TD-F-113-012&plan_year=93&projkey=PG9303-1059&target=plan&highStr=*&check=0&pnchDesc=%E8%81%9A%E5%90%88%E8%84%A2%E9%8F%88%E5%8F%8D%E6%87%89%E6%8A%80%E8%A1%93%E6%96%BC%E5%9F%BA%E5%9B%A0%E6%94%B9%E9%80%A0%E7%8E%89%E7%B1%B3%E4%B9%8B%E5%8D%8A%E5%AE%9A%E9%87%8F%E5%8F%8A%E5%93%81%E7%B3%BB%E9%91%91%E5%AE%9A%E6%96%B9%E6%B3%95%E4%B9%8B%E9%96%8B%E7%99%BC%E8%88%87%E6%AA%A2%E6%B8%AC
Appears in Collections:食品暨應用生物科技學系

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