基于巰基識別的DNA、蛋白質(zhì)生物傳感器的研制.pdf

1、中南大學(xué)碩士學(xué)位論文基于巰基識別的DNA、蛋白質(zhì)生物傳感器的研制姓名：李凱申請學(xué)位級別：碩士專業(yè)：應(yīng)用化學(xué)指導(dǎo)教師：王建秀劉又年20080529碩士學(xué)位論文ABSTRACTABSTRACTFluorescencespectroscopyandvoltammetryhavebeenutilizedtothedeterminationofultratracelevelsofbiomoleculessuchaspeptidesproteins

2、andDNApreimmobilizedontotheelectrodesurfaceTheintroductionofthefunctionalthi01specificreagentsinthesemethodsincreasesthesensitivityselectivityandtheabilitytorecognizethespeciesanalyzedThemaincontentsofthethesisareasfollo

3、ws：1Todevelopamethodforthedetectionofsurfaceconfinedpeptidescontainingcysteineresiduesoroligodeoxynucleotides(ODNs)whose3’endsmodifiedwiththiolgroups，athiolspecificfluorescentcrosslinkerN一(9一acridinyl)maleimide(NAM)wasus

4、edThepeptidesstudiedhereinincludeboththeoxidizedandreducedformsofglutathione，andahexapeptide(FT)Peptidesarefirstattachedontotheactivated11一mercaptoundecanoicacid(MUA)terminatedalkanethiolselfassembledmonolayers(，SAMs)and

5、thenderivatizedwimN√氣MThecysteineresidueswasdeterminedviacombinationofelectrochemicaldesorptionwithfluorescencedetectionGSHconcentrationaslowas40pmolL～CanbemeasuredThefluorescenceintensityinthecaseofFTiSabout3timesashigh

6、asthatforGSH，whichiSconsistentwiththemolarratioofcysteineresiduesinthesetwomoleculesTheanalyticalperformanceofgeneanalysiswasalsoevaluatedthroughtheanalysesofacomplementarytargetandtargetswithvaryingnumbersofmismatchingb

7、asesThemethoddescribedhereiSsimple，sensitive，reproducible，anddoesnotrequiresophisticatedanalyticalinstrumentationandseparationprocedures2DevelopmentofsensitiveandselectivemethodstodetectproteinattracelevelsiSofgreatbiolo

8、gicalimportanceConsensusdoublestrandedoligonucleotide(dsDNA)wasfirstformedontogoldelectrodesviahybridizationofthiolatedsinglestrandedDNA(ss—DNA)／hexanethiol(HT)mixedselfassembledmonolayerwiththecomplementaryDNAtocapturew