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ID:33505658
大小:2.37 MB
页数:56页
时间:2019-02-26
《内毒素休克小鼠肝脏线粒体差异蛋白质组的鉴定》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、硕士学位论文before2-Delectrophoresis,andthenseparateuptothreedifferentproteinsamplesatthesametimeinonetwo.dimensionalgel.Theapplicationoftheinternalstandardcouldfurtherincreasethecredibilityoftheexperiment,andensuretheresultscouldreflecttherealbiologicaldifferences,
2、whileavoidinfluenceofsystematicerrorsonexperimentalresults.SinceThemostobviousadvantageofDIGEsystemisintegratingtheadvantagesofbothCyDyeDIGEdyemultiplelabelingmethodandDeCyderdifference2-Danalysissoftware.DeCydersoftwaretakest11eadvantageofthespotsCO·detecting
3、algorithm,whichCanautomaticallydetectedfluorescenceimages,eliminatebackground,quantifiy,normalizeandmatchspotsingel,thus,systematicerrorscausedbydifferentoperatorsconbeeliminated.BasedontheadvantagesofDIGEtechnologyplatform,greatopportunitiescomestoustodiscove
4、rtheinformationofdifferentialproteomecontainedinmicelivermitochondria、析tllendotoxicshockandprovidenoveltheoriesforsepsisresearch.Atechnologyplatformbasedondifferencegelelectrophoresis,MALDITOF/TOFmassspectrometryandbioinformaticsanalysiswasestablished缶st,andth
5、ensomeassaysonitssensitivityandreproducibilitywasdoneinourresearch.AfterduplicatingtheendotoxicshockmodelofBALB/cmiceafteranesthesia,wecollectedmiceliver,applieddifferentialcentrifugationanddensitygradientcentrifugationmethodtoextractpurifiedlivermitochondfia,
6、andextractedtheentiremitochondrialproteinsbylysisbuffer.ThenwegottheexpressionpatternsoflivermitochondrialdifferentialproteomeofendotoxicshockmicebyusingDIGEtechnology.ThroughanalysisbyuseofDeCyderdifference2一Danalysissoftware,26differentialproteinspotswerefou
7、nd.,nlenumbersofup—regulatedanddown-regulatedproteinsareboth13.Finally,atotalof22proteinshavebeenidentifiedamongthesedifferentialproteinspotsbyusingmatrix-assistedlaserdesorptionionizationmassspectrometry(MALDI—TOF/TOF).neCO.expressionofthesedifferentialprotei
8、nsindicatesthattheymusthavedirectorindirectrelations.Therefore,howtofindoutanddiscoverthepathophysiologymeaningoftheserelationsinendotoxicshocklivermitochondriaisthekeyforfurtherre
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