生物分析检测技术.ppt

Intransgenicplants,transgenecopynumbercangreatlyinfluencetheexpressionlevelandgeneticstabilityofthetargetgene,makingestimationoftransgenecopynumberanimportantareaofgeneticallymodified(GM)cropresearch.TransgenecopynumbersarecurrentlyestimatedbySouthernanalysis,whichislaboriousandtime-consuming,requiresrelativelylargeamountsofplantmaterialsandmayinvolvehazardousradioisotopes.第96页,共103页，星期六，2024年，5月Reportherethedevelopmentofasensitive,high-throughputreal-time(RT)-PCRtechniqueforestimatingtransgenecopynumberinGMrice.UseTaqManquantitativeRT-PCRandcomparisontoanovelriceendogenousreferencegenecodingforsucrosephosphatesynthase(SPS)todeterminethecopynumbersoftheexogenousbeta-glucuronidase(GUS)andhygromycinphosphotransferase(HPT)genesintransgenicrice.第97页,共103页，星期六，2024年，5月ThecopynumbersoftheGUSandHPTinprimaryricetransformants(T0)werecalculatedbycomparingquantitativePCRresultsoftheGUSandHPTgeneswiththoseoftheinternalstandard,SPS.WithoptimizedPCRconditions,achievedsignificantlyaccurateestimatesofone,two,threeandfourtransgenecopiesintheT0transformants.CopynumberestimationsofboththeGUSreportergeneandtheHPTselectivemarkergeneshowedthatrearrangementsoftheT-DNAoccurredmorefrequentlythanisgenerallybelievedintransgenicrice.

第98页,共103页，星期六，2024年，5月AnexpressionanalysisofagenefamilyencodingplasmamembraneaquaporinsinresponsetoabioticstressesinArabidopsisthaliana.

JangJY,KimDG,KimYO,KimJS,KangH.

DivisionofAppliedPlantScienceandAgriculturalPlantStressResearchCenter,CollegeofAgricultureandLifeSciences,ChonnamNationalUniversity,300Yongbong-dong,Buk-gu,Gwangju500-757,Korea.

PlantMolBiol.2004Mar;54(5):713-25.第99页,共103页，星期六，2024年，5月Aquaporinbelongstoahighlyconservedgroupofmembraneproteinscalledmajorintrinsic