18808.x染色體調控元件的定位及dna親和純化技術的探索

1、南開大學碩士學位論文X染色體調控元件的定位及DNA親和純化技術的探索姓名：郭宏超申請學位級別：碩士專業(yè)：細胞生物學指導教師：陳凌懿201205AbstractAbstractXchromosomeinactivation(XCI)isthemechanismofbalancingtheexpressionofXlinkedgenesbetweenmaleandfemaleinmammalsItincludestwotypesofinac

2、tivation：imprintedXCI(PaternalXchromosome(Xp)isalwaysinactivated)andrandomXCI(PaternalXchromosomeandmatemalXchromosome(XpliSrandomlyinactivated)HoweverXcontrollingelement(Xce)andepigeneticfactorswillskewtheinactivationra

3、tebetweenXpandXm(skewingtowardXporXm)Throughcomparingtheactivitiesof居扔majorpromo胞r，Xite，DXPas34，Xistminimalpromo跑rbetweenMusmusculusmusculus(129)andMusmusculuscastaneusstrain(cast)byLuciferaseAssayandElectrophoreticMobil

4、ityShiftAssay(EMSA)，wefoundthatXistminimalpromo紀risaXce，andthe一88sitemutation(A／C)isthemainreasonofthedifferentXisttranscriptionalactivitybetweenX129andX。8吼Inaddition，weinitiallyestablishedtheexperimentalsystemofDNAaffin

5、itychromatographyWithaDNAprobefromXistintronJ『，PTBWaspurifiedandidentifiedwithmassspectrometryHoweverEMSASupershiftandChromatinImmunoprecipitation(ChIP)resultsindicatedthatPTBdoesnotbindtothisDNAprobeInordertofindthetarg

6、etprotein，weneedtoimproveourDNAaffinitychromato鏟aphXincludingthepurificationefficiencytherecoveryefficiencyoftargetproteinandtheresolutionofSDS—PAGEgelandSOonKeywords：SkewedXCI，Xce，Xistminimalpromoter，DNAaffinitychromato