Global Epigenetic Changes Induced by SWI2SNF2 Inhibitors Characterize Neomycin-Resistant Mammalian Cells 英文参考文献.docVIP

下载本文档

4
0
约5.69万字
约 13页
2017-05-11 发布于上海
举报
版权申诉

Global Epigenetic Changes Induced by SWI2SNF2 Inhibitors Characterize Neomycin-Resistant Mammalian Cells 英文参考文献.doc

1、本文档共13页，可阅读全部内容。
2、原创力文档（book118）网站文档一经付费（服务费），不意味着购买了该文档的版权，仅供个人/单位学习、研究之用，不得用于商业用途，未经授权，严禁复制、发行、汇编、翻译或者网络传播等，侵权必究。
3、本站所有内容均由合作方或网友上传，本站不对文档的完整性、权威性及其观点立场正确性做任何保证或承诺！文档内容仅供研究参考，付费前请自行鉴别。如您付费，意味着您自己接受本站规则且自行承担风险，本站不退款、不进行额外附加服务；查看《如何避免下载的几个坑》。如果您已付费下载过本站文档，您可以点击这里二次下载。
4、如文档侵犯商业秘密、侵犯著作权、侵犯人身权等，请点击“版权申诉”（推荐），也可以打举报电话：400-050-0827(电话支持时间：9:00-18:30)。
5、该文档为VIP文档，如果想要下载，成为VIP会员后，下载免费。
6、成为VIP后，下载本文档将扣除1次下载权益。下载后，不支持退款、换文档。如有疑问请联系我们。
7、成为VIP后，您将拥有八大权益，权益包括：VIP文档下载权益、阅读免打扰、文档格式转换、高级专利检索、专属身份标志、高级客服、多端互通、版权登记。
8、VIP文档为合作方或网友上传，每下载1次，网站将根据用户上传文档的质量评分、类型等，对文档贡献者给予高额补贴、流量扶持。如果你也想贡献VIP文档。上传文档

Global Epigenetic Changes Induced by SWI2SNF2 Inhibitors Characterize Neomycin-Resistant Mammalian Cells 英文参考文献

GlobalEpigeneticChangesInducedbySWI2/SNF2 InhibitorsCharacterizeNeomycin-ResistantMammalian Cells PopyDutta1.¤,GoutamKumarTanti1.,SoniSharma1.,ShyamalK.Goswami1,SnehaSudhaKomath1, MartyW.Mayo2,JoelW.Hockensmith2*,RohiniMuthuswami1* 1SchoolofLifeSciences,JawaharlalNehruUniversity,NewDelhi,Delhi,India,2DepartmentofBiochemistryandMolecularGenetics,UniversityofVirginia,Charlottesville, Virginia,UnitedStatesofAmerica Abstract Background:Previously,weshowedthataminoglycosidephosphotransferasescatalyzetheformationofaspecificinhibitor of the SWI2/SNF2 proteins. Aminoglycoside phosphotransferases, for example neomycin-resistant genes, are used extensivelyasselectionmarkersinmammaliantransfectionsaswellasintransgenicstudies.However,introductionofthe neomycin-resistantgeneisfraughtwithvariabilityingeneexpression.Wehypothesizedthattheintroductionofneomycin- resistant genes into mammalian cells results in inactivation of SWI2/SNF2 proteins thereby leading to global epigenetic changes. Methodology:Usingfluorescencespectroscopywehaveshownthattheinhibitor,knownasActiveDNA-dependentATPase ADomaininhibitor(ADAADi),bindstotheSWI2/SNF2proteinsintheabsenceaswellaspresenceofATPandDNA.This bindingoccursviaaspecificregionknownasMotifIaleadingtoaconformationalchangeintheSWI2/SNF2proteinsthat precludesATPhydrolysis.ADAADiisproducedfromaplethoraofaminoglycosidesincludingG418andStreptomycin,two commonly used antibiotics in mammalian cell cultures. Mammalian cells are sensitive to ADAADi; however, cells stably transfectedwithneomycin-resistantgenesarerefractorytoADAADi.Inresistantcells,endogenousSWI2/SNF2proteinsare inactivatedwhichresultsinalteredhistonemodifications.Microarraydatashowsthatthechangesintheepigenomeare reflected in altered gene expression. The microarray data was validated using real-time PCR. Finally, we show that the epigeneticchangesarequantized. Significance:Theuseofneomycin-resistantgenesrevolutionizedmammaliantransfectionseventhoughquestionslinger aboutefficacy.Inthisstu