(53(1):9-17)Developing the Techniques for Transgenic Crops Detection -Detection of Genetically Modified Soybeans and Maize by 35S Promoter Molecular Marker

本研究係分別以轉抗殺草劑基因之大豆以及轉抗蟲基因之玉米作為模式植物，建立基因轉殖植物在DNA層次之初步檢測分析。研究中基因轉殖與非基因轉殖之玉米及大豆共十七個樣品，經比較不同DNA萃取成本、DNA純度以及聚合酵素連鎖反應結果，以 WizardTM Genomic DNA Purification Kit 萃取結果較佳。另比較利用巢式聚合酵素連鎖反應 (Nested Polymerase Chain Reaction) 以及一般聚合酵素連鎖反應，進行基因轉殖大豆及玉米之鑑定，另以歐盟認可之基因轉植產品偵測試劑組 (Biosmart Allin 1.0 GMO Screening System) 為對照，經依作物調整其聚合酵素連鎖反應條件，結果可正確測得基因轉殖玉米及大豆樣品中的CAMV 355 啟動子特有之150bp DNA片段；此外，依CAMV 35S啟動子序列，所合成之35S 1 / 35S 2 引子以及35S-F / 35S-R引子，亦可分別用以測得基因轉殖樣品中CAMV 35S啟動子特有之195bp DNA片段與207bp之DNA片段。Biotechnology has enabled the modification of crops in a very precise way thereby improving productivity and yield. Till 2003 about 78 transgenic crops were through safety evaluated and already approved. Since that, the problem for discriminating between transgenic crops and non-transgenic crops in the market was posed. In this study, we used herbicide resistance transgenic soybean and Bacilnus tumufacisos (Bt) corn as model plants for setup the DNA level detection of transgenic crops. Three commercial DNA extraction kits were evaluated for DNA quality and PCR efficiency. The results showed that the Wizard® Genomic DNA Purification Kit can be manipulated easily to obtain DNA with good quality. The results of transgenic crops detection revealed that Nested PCR (Nest Polymerase Chain Reaction) analysis based on Biosmart Allin 1.0 GMO Screening System allowed the detection of transgenic soybean and transgenic corn by CaMV 35S promoter specific gene fragment of 150bp. Furthermore, the synthesized primers of 35S 1 / 35S 2 and 35S-F/35S-R also showed very efficient in detected the CaMV 35S promoter and produced the fragments of 195bp and 207bp respectively