多重实时荧光PCRTaqMan-MGB杂交探针标记法快速鉴定转基因玉米MIR162王凤军，许海锋，陈强，杨伊平，金浩然（浙江经贸职业技术学院,浙江杭州310018）摘要：为对转基因玉米MIR162的进口实行监测、规范转基因玉米在国内市场中的流通，建立一种转基因玉米MIR162准确快速的检测方法，通过设计转基因玉米MIR162品系特异性序列的引物和探针，摸索和优化多重荧光PCR体系和参数，开发建立多重实时荧光PCRTaqMan-MGB杂交探针标记法快速鉴定转基因玉米MIR162。结果表明，转基因玉米MIR162品系标准品和平行样品中内标基因和品系特异性基因均出现明显扩增曲线，其他转基因玉米品系标准品仅内标基因有扩增曲线，空白对照无扩增曲线，说明该TaqMan-MGB探针对转基因玉米MIR162品系具有扩增特异性。该方法可作为鉴定筛查转基因玉米MIR162品系及其转化体成分的有效方法，用于规范管理转基因产品在市场上的流通。关键词：转基因玉米MIR162；多重实时荧光PCR；TaqMan-MGB杂交探针标记法；快速鉴定RapidIdentificationofTransgenicMaizeMIR162byMultipleReal-TimeFluorescentPCRwithTaqMan-MGBHybridizationProbeLabelingWANGFeng-jun,XUHai-feng,CHENQiang,YANGYi-ping,JINHao-ran（ZhejiangInstituteofEconomicandTrade,Hangzhou310018,China）Abstract:InordertomonitortheimportoftransgenicmaizeMIR162andstandardizethecirculationoftransgenicmaizeindomesticmarket,anaccurateandfastdetectionmethodfortransgenicmaizeMIR162wasestablished.BydesigningprimersandprobesforthespecificsequenceoftransgenicmaizeMIR162,themultiplefluorescentPCRsystemandparameterswereexploredandoptimized,andamultiplereal-timefluorescentPCRwithTaqMan-MGBhybridizationprobelabelingmethodforrapidlyidentifyingtransgenicmaizeMIR162wasdevelopedandestablished.Theresultsshowedthattheinternalstandardgeneandline-specificgeneoftransgenicmaizeMIR162linestandardandparallelsamplespresentedobviousamplificationcurves,andonlytheinternalstandardgeneofothertransgenicmaizelinestandardsexhibitedamplificationcurves,whiletheblankcontrolhadnoamplificationcurves,indicatingthattheTaqMan-MGBprobehadamplificationspecificityfortransgenicmaizeMIR162strain.ThismethodcanbeusedasaneffectivemethodtoidentifyandscreentransgenicmaizeMIR162strainsandtr...
