水稻内标准基因检测方法国际协同验证及四种转基因棉花品系特异性检测方法研究

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1、上海交通大学硕士学位论文AbstractINTERNATIONALVALIDATIONOFRICEENDOGENOUSREFERENCEGENEDETECTIONSYSTEMSANDDEVELOPMENTOFPCRDETECTIONMETHODSOFFOURVARIETIESGENETICALLYMODIFIEDCOTTONSAbstractToensuretheimplementofgeneticallymodifiedorganisms(GMO)labelingregulationintheworldandperfectthedetectionm

2、ethodssystemforGMOs,twoaspectsthatrelatedtoGMOsidentificationandquantificationarecoveredinthisstudy.1.TheinternationalvalidationofSPSgeneasriceendogenousreferencegeneforqualitativeandquantitativePCRanalysis.EndogenousreferencegeneisveryimportantforGMOsdetectionandquantification

3、.Onerice(Oryzasativa)gene,SucrosePhosphateSynthase(SPS),hadbeenprovedtobesuitablefortheendogenousreferenceofgeneticallymodified(GM)ricedetectionandquantificationinourpreviousstudy.HereinweinvitedtwelveGMOdetectionlaboratoriestoparticipateinthecollaborativeringtrialforinternatio

4、nalvalidationoftheIV上海交通大学硕士学位论文AbstractSPSgeneasriceendogenousreferencegene.InthecollaborativeringtrialofSPSgenevalidation,thesubmittedresultsfromtwelvelaboratoriesconfirmedthespeciesspecificity,lessallelicvariationandstablesinglecopynumberoftheSPSgene.Furthermore,limitofdetec

5、tion(LOD)ofSPSqualitativePCRwasprovedtobe0.1%,andlimitofquantification(LOQ)forquantitativePCRsystemwasatleast23copieshaploidricegenomicDNAwithhighPCRefficiencyandlinearity.Additionally，thebiasbetweenthetestandtruevaluesofeightblindsamplesrangedfrom1.%to26.52%.Allthevalidatedres

6、ultsindicatedthattheSPSgeneissuitableforusingasendogenousreferencegeneforGMriceanditsderivatesidentificationandquantification.2.QualitativeandquantitativePCRmethodsforevent-specificdetectionofGMcotton.Event-specificPCRisbecomingthemosteffectiveandimportantmethodforGMOdetectiona

7、ndquantification.Inthisstudy,the5’-integrationjunctionsequencebetweenhostplantDNAandtheintegratedgeneconstructofMon15985andthe3’-integrationjunctionsequenceofMon88913wererevealed.SpecificPCRprimersandTaqManprobesweredesignedbasedupontherevealedsequenceandtheevent-specificqualit

8、ativeandquantitativedetectionmethodswerefirstlyestabli