单细胞凝胶电泳检测DNA损伤的最新研究进展

20112561230～1234JournalofNuclearAgriculturalSciences2011-02-172011-04-14310712971986-、。[e-mail]1975-、。[e-mail]1000-8551201106-1230-05DNA/310018DNA、DNA。DNA、、DNA、、。、DNA。。。DNATHELATESTPROGRESSINSINGLECELLGELELECTROPHORESISSCGEBASEDONDNADAMAGEDETECTIONZHANGChun-hongLINXin-daCollegeofLifeSciencesZhejiangProvincialKeyLaboratoryofBiometrologyandInspection＆QuarantineChinaJiliangUniversityHangzhouZhejiang310018AbstractDNAdamagedetectioncandetectDNAdamagecausedbythepesticideandirradiation．WiththeincreasingdemandsofDNAdamagedetectionthedevelopmentofarapidhighthroughputandstraightforwardDNAdamagedetectingtechniquehascriticalbiologicalsignificanceforSingleCellGelElectrophoresisSCGEisastraightandaccuratewaytodetecttheDNAdamage．InrecentyearsthethroughputandaccuracyofthedetectionSCGEmethodhavebeenimprovedsignificantlybyapplyingnewmaterialsandnewtechnologies．ThispaperreviewedthemostrecentlyreportedSCGEbasedDNAdamagedetectiontechnique-microwellarraymethodandconventionalSCGEmethodandtheprospectwerealsodiscussed．KeywordsDNAdamagesinglecellgelelectrophoresismicrowellarrayDNA。DNA1DNA2。SCGEDNA。71%DNA37fanconianemia、ataxiatelangiectasiadownsyndrome8～14DNA03216DNADNA1213DNA。DNADNADNA。DNA。1DNADNA、、、、、。DNADNA32P--32P-postlabeling-TLCassay、ISBassay、--GC-MS-SIM、--HPLC-GC-MS、-HPLC-ECD、。。GC-MSHPLC-ECDDNADNATUNEL、LM-PCRligation-mediatedPCR、RD-PCRrandomizedterminallinker-dependentPCR、RT-PCRrealtimePCRGFP、γH2AX、SCE、DNAUDS、dUTPDNA、、、、、。1.1singlecellgelelectrophoresisSCGEDNADNA14。cometassay。Ostling15Singh16～18DNA。200DNA。SCGE、、、、DNA-DNA、DNA-、、、、。SCGE、、、、、、。pH=8pH＞13152。。、RNA、DNA1。DNADNA。Tris。、、Olive、、、/DNA、、19。1.2GelbondMcNamee20SCGEGelbond。GelbondGelbond。GelbondLab-TekII1、2、4、816。Lab-TekIIGelbond。Lab-TekIIGelbondSCGE1。GelbondSCGE。1.3DavidSCGEGelbond、、DNA、、132125、、、、microwellarraySCGE21～31。1SCGE、GelbondFig．1BasicflowofSCGEGelbondarrayandmicrowellarraymethods1SU-8SU-82000SU-8UV。SU-820～50μm。。Gelbond4℃。SCGE、、、、NIS-ElementsNikonMATLAB、、/3。SCGEDNA。2SCGE、GelbondSCGE、Gelbond31。SCGE。GelbondSCGESCGE23216DNA1SCGE、GelbondTable1ComparisonoftheSCGEGelbondarrayandmicrowellarraymethodsSCGEGelbondGelbondarraymicrowellarraymethodbottomglueGelbondGelbondsampletreatmentSCGEthroughput1696sensitivitydataacquisitionSCGEresultanalysisSCGErepeatabilitySCGE。SCGE、SCGEGelbond8.8%SCGE21。3、、、DNA、、。SCGEDNA。、。SCGEDNA。SCGE。DNASCGEDNA。1BlanpainCMohrinMSotiropoulouPAPassegueE．DNA-damageresponseintissue-specificandcancerstemcellsJ．CellStemCell2011816－292WoodruffRCThompsonJNJ．TheroleofsomaticandgermlinemutationsinagingandamutationinteractionmodelofagingJ．JournalofAnti-AgingMedicine2003629－393．J．2005192159－1624YinXHLiSNZhangLZhuGNZhuangHS．EvaluationofDNAdamageinChinesetoadBufobufogargarizansafterinvivo3321JournalofNuclearAgriculturalSciences20112561230～1234exposuretosublethalconcentrationsoffourherbicidesusingthecometassayJ．Ecotoxicology2008174280－2865．J．2008222196－1996OstlingOJohansonKJ．Microelectrophoreticstudyofradiation-inducedDNAdamagesinindividualmammaliancellsJ．BiochemicalandBiophysicalResearchCommunications19841231291－2987HifzurRSAnuragSSubashCGRameshCMAlokDDayaKSDebapratimKC．DNADamageinducedbyindustrialsolidwasteleachatesinDrosophilamelanogasteramechanisticapproachJ．EnvironmentalandMolecularMutagenesis200849206－2168BarbaraCGWouterWWQuintenWAnnetteLMFreAHansJMalgorzataZZ．InducibilityofnuclearRad51fociafterDNAdamagedistinguishesallFanconianemiacomplementationgroupsfromD1/BRCA2J．MutationResearch200659439－489LowGKFokEDTingAPHandeMP．OxidativedamageinducedgenotoxiceffectsinhumanbroblastsfromXerodermaPigmentosumgroupapatientsJ．TheInternationalJournalofBiochemistry＆CellBiology2008402583－259510ElenaVMathiasT．MolecularmechanismsinrenaldegenerativediseaseJ．SeminarsinCell＆DevelopmentalBiology201021831－83711RongGDongYXWeiDW．IdentificationandanalysisofnewproteinsinvolvedintheDNAdamageresponsenetworkofFanconianemiaandBloomsyndromeJ．Methods20094872－7912AndreaBRobertaMSilviaF．l-CarnitineenhancesresistancetooxidativestressbyreducingDNAdamageinataxiatelangiectasiacellsJ．MutationResearch2008650165－17413DimitriosPNikiMNMariaZAndreasFSmaragdiTStylianosMP．DNAdamageandrepairefficiencyinymphocytesfromschizophrenicpatientsJ．CancerLetters200420433－4014ZbigniewMKatarzynaJMichalKTomaszSJerzySAlinaMBAnnaSJanuszB．DNAdamageandrepairinchildrenwithDown’ssyndromeJ．MutationResearch2008637118－12315ZhangLWangJZhuGNSuL．PubertalexposuretothiodiazolecopperinhibitsthyroidfunctioninjuvenilefemaleratsJ．ExperimentalandToxicologicalPathology2010622163－17116XingMLJianJWJianMX．CharacterizingtheriskassessmentofheavymetalsandsamplinguncertaintyanalysisinpaddyfieldbygeostatisticsandGISJ．EnvironmentalPollution20061412257－26417NarendraPSMichaelTMRaymondRTEdwardLS．AsimpletechniqueforquantitationoflowlevelsofDNAdamageinindividualcellsJ．ExperimentCellResearch1988175184－19118NarendraPSRaymondRTRalphESEdwardLS．AmicrogelelectrophoresistechniqueforthedirectquantitationofDNAdamageandrepairinindividualfibroblastsculturedonmicroscopeslidesJ．Mutationresearch1991252289