植物转基因成分PCR检测内对照系统的建立

为了建立适用于多种植物的转基因成分PCR检测的内对照系统，本文针对植物叶绿体DNArbcL基因的保守区域，设计了一对扩增片段为433bp的PCR引物。通过对23种植物的PCR扩增表明，该引物不但在4种单子叶植物（大米，玉米，小麦，洋葱），10种原始花被亚纲的双子叶植物（甘蓝，白菜，大豆，豇豆，花生，胡萝卜，芹菜，菠菜，大麻，棉花）及7种合瓣花亚纲的双子叶植物（圣女果，番茄，辣椒，马铃薯，南瓜，黄瓜，菊苣）中得到了稳定一致的扩增结果，而且在低等的藻类植物（海,经须菜）中也得到了特异性的扩增结果。进一步对扩增片段进行的DNA序列测定与分析表明，扩增片段的变异水平较低，具有较高的保守性。本系统的建立有助于排除PCR检测时的假阴性结果，从而提高检测的准确性，而且能克服现行的“一种植物一种检测内对照”的弊端，有利于提高检测效率，缩短检测周期。

Establishment of A Universal Inner Positive Control for PCR Detection of Genetically Modified Ingredient in Plants

In the present study,a primer pair for the amplification of rbcL 433bp fragment was designed.In order to find out whether this primer pair can be used as inner positive control for PCR detection of genetically modified ingredient in most plants,it was used to amplify DNA from 23 various plant species including monocotyledon ( Allium cepa,Triticum aestivum,Zea mays,Oryzasativa ), dicotyledon in Archichlamydeae ( Brassica oleracea,Brassica pekinensis,Glycine max,Vigna unguiculata,Arachis hypogaea,Daucus carota,Apium graveolens, Spinacia oleracea,Cannabis sativa,Gossypium hirsutum ),dicotyledon in Sympetalae ( Lycopersicon esculentum,Lycopersicon esculentum,Capsicum frutescens,Solanum tuberosum,Cucurbita moschate,Cucumis sativusl,Cichorium intybus ) and algae( Laminaria angustata,Gracilaria lemaneiformis ).The results showed that the primer pair worked very well over all the samples,and the specific fragment could be amplified via this primer pair over wide annealing temperature.The validity of amplified fragment was further tested via DNA sequencing.These results suggest that amplification of this gene fragment via this primer pair can be used as inner positive control for PCR detection of genetically modified ingredient in most plants, which will be helpful to the improvement of detection accuracy,as well as the reduction of detection period.