农业转基因生物检测技术的初步研究
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摘要
自1983年首例转基因作物(Genetically Modified Crops,GMC)问世以来,作为一项科学新产物,在给人类带来巨大恩惠的同时也可能带来了潜在的风险。随着转基因原料及食品大量涌向市场,其生物安全性(Biosafety)问题便受到了广泛争论,并引发了政治、贸易乃至道德伦理上的各种争端。世界各国纷纷出台各种管理政策和法规。我国规定从2002年3月20日起,凡在中国境内销售的大豆、玉米等及其制品,若属转基因产品必须进行标识。由于食品标识以及食品制造商、销售商和消费者等多方面的需要,将转基因食品与常规食品区分开,迫切需要一套切实可行的检测方法以满足需要。
     本研究初步建立了转基因农作物常用的调控基因35S启动子和nos终止子的PCR检测技术,检测结果表明:本实验的转基因检测下限为0.5%(w/w),即当受检材料中的转基因成分占非转基因成分等于或大于0.5%时可以检出。并将35S启动子和nos终止子用地高辛标记后制备成基因探针,建立了转基因检测的分子杂交技术—点杂交。杂交试验证明PCR扩增的35S启动子和nos终止子为特异性扩增,当材料转基因含量降低时,检测信号则明显降低。此项检测技术具有高特异性但灵敏度低,不适合市场上的转基因检测。
The first Genetically Modified Crops (GMC), which was born in the 1983 maybe have brought up potential risk when it gave human much favor. As foods produced from Genetically Modified organisms (GMOs) or food products containing GMOs are appearing more and more frequently on the market, the problem of biosafety is being extensively debated and resulted kinds of conflict on politics , trade and even religion. As a result, many countries promulgated laws and regulation on GMO biosafety. Our government required the agricultural products marketed in china such as soybean, maize , their products and so on ,if it produced from GMOs or contained GMOs, must be labeled before flowed into market A feasible detection method should be built to meet the needs from foods labelling, producer and consumers.
    In this study, a PCR detection method on CaMV 35S promoter and nos terminator which are extensively used in GMC as regulatory elements. The PCR detection result showed: the practical detection limit was 0.5%(w/w), which meant this method could detect the GM soybean when its weight proportion was equal or exceeded 0.5%. Another detection method, dot blotting, was built also based on 35S promoter and nos terminator. The two genes were labeled by digoxin to done as probe used in dot blotting. The blotting detection showed: the PCR detection on 35S and nos is correct, while when the GMS content decrease the detection signal weakened. So this method was not fit to be applied in market detection.
引文
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