| Summary: | 碩士 === 中興大學 === 食品暨應用生物科技學系 === 95 === In recent years, a number of genetically modified (GM) crops have been developed and commercialized by many agricultural biotechnological companies. This has been accompanied by a considerable increase in the diversity of genetically modified organisms (GMO) currently approved worldwide. Governments in many countries have established a policy of labeling all food and feed containing or produced by GMOs. In Taiwan, food products that contained more than 5% GM are required to put on the label. The purpose of the labeling system is to inform the consumers of the presence of GMOs in the product, thus providing the consumers with the option to select their preferred products.
In this study, we developed a method based on semi-quantitative PCR to detect amylopullulanase (APU) transgenic rice. We expect the system could qualitated and quantitated GM content in food simultaneously . We designed the endogenous primers Osps-1F/Osps-1R (sucrose phosphate synthase gene) and exogenous primers Apu-2F/Apu-2R (APU gene) , which yielded PCR products of 213 bp and 163 bp, respectively. The primers had good specificity in selecting transgenic rice. In simplex and duplex PCR detection systems, the detection limit of the systems were 0.1%, and according to the concentration of GM and the ratio of brightness of the correlation coefficient of semi-quantitative standard curve were 0.9767 and 0.9957, respectively. It showed that duplex PCR system was more accurate.
The real-time PCR used SYBR GreenⅠsystem as control group to compare semi-quantitative PCR system. When detecting a lot of samples, semi-quantitative PCR system could separate them from more than 7% or less than 3% to decrease the numbers of samples for quantitating by real-time PCR. Analysis of rice endogenous gene by duplex PCR with Osps-1F/Osps-1R and Apu-2F/ Apu-2R primers in processed food, we found that DNA would breakdown by processing steps. In rice transgene, we obtained no samples contained APU gene. It showed that non-legalized APU transgenic rice has not sold in the market.
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