27989.耐mtx小鼠nih3t3細(xì)胞中dms的分子結(jié)構(gòu)研究

1、哈爾濱醫(yī)科大學(xué)博士學(xué)位論文耐MTX小鼠NIH3T3細(xì)胞中DMs的分子結(jié)構(gòu)研究姓名：李明申請學(xué)位級別：博士專業(yè)：遺傳學(xué)指導(dǎo)教師：傅松濱20100501哈爾濱醫(yī)科人學(xué)博l：學(xué)位論文ArchitecturesofDMsiUMTXresistantNIH3T3CellsatSequence—levelResolutionAbstractObjeetive：Doubleminutes(DMs)arecytogenetichallmarkbeari

2、ngextrachromosomallyamplifiedgenesincancercellsMethotrexate(MTX)resistantMousefibroblastlinesNIH3T3canmaintainDMsthatcontainDhfr(dihydrofolatereductase)geneDhfrisknowntelatetoMTXresistantMethod：First，wegotthegenome—widec

3、opynumberalterationofMTXresistantcellsbyarray—CGHSecond，theexpressionlevelsofgeneswithhighercopynumberweredetectedbyRealtimeRTPCRandWesternblotThen，integratedanalysisofexpressionandarrayCGHdatawasdonetofindthefunctionoft

4、hegeneswithhighercopynumberFollowing，DMswasseparatedbymicro—dissectionand、amplifiedbyDOP—PCR，andsequencesofDMswereidentifiedby454sequencingResults：AmpliconsofDMsarefrom5A1、10D2and13C3，includingPexl，Gatadl，4930511M11Rik，A

5、nkibl，Kritl，4932412H11Rik，MterfAkap9，Mdm2，Slc35e3，Nupl07，Rasg吐Msh3，DhfrAnkrd34band24930405M20RikTheregionsofamplificationareabout10MbandnearofthefragilesitesTheexpressionofGatadl，Ankibl，Kritl，MterfMdm2，S1c35e3，Msh3andDhf

6、rincreasedinMTXresistantcellsIntheampliconsofDMs，thegenedensityandGCpercentarehigherandtherepeatpercentislessthanthatofthemousegenomeLesshomologusrecombineationandlotsofnon—homologusrecombinationwerefoundinDMsConclusion：

7、DMsgenerationispossiblelyamulti—stepandmulti—eventprocessNonhomologousendjoiningmechanismwasindicatedtoplaytheleadingrolefortheDNAbreakpointsrejoiningduringDMsformationKeywords：MTX；DMs；non—homologusrecombination；drugresi