国产三级在线看完整版-内射白嫩大屁股在线播放91-欧美精品国产精品综合-国产精品视频网站一区-一二三四在线观看视频韩国-国产不卡国产不卡国产精品不卡-日本岛国一区二区三区四区-成年人免费在线看片网站-熟女少妇一区二区三区四区

儀器網(wǎng)(yiqi.com)歡迎您!

| 注冊2 登錄
網(wǎng)站首頁-資訊-話題-產(chǎn)品-評測-品牌庫-供應(yīng)商-展會-招標(biāo)-采購-知識-技術(shù)-社區(qū)-資料-方案-產(chǎn)品庫-視頻

資料庫

儀器網(wǎng)> 資料庫>酵母單雜交系統(tǒng)Clontech pHis2說明書

酵母單雜交系統(tǒng)Clontech pHis2說明書

本文由 上海起發(fā)實(shí)驗(yàn)試劑有限公司 整理匯編

2018-09-29 10:03 6437閱讀次數(shù)

掃    碼    分   享
文檔僅可預(yù)覽首頁內(nèi)容,請下載后查看全文信息!
立即下載
酵母單雜交系統(tǒng)ClontechpHis2說明書DescriptionpHIS2isareportervectorthatcanbeusedinyeastone-hybridassaystoidentifyandcharacterizeDNA-bindingproteins.ThevectorwasspecificallydesignedforusewiththeBDMatchmakerOne-HybridLibraryConstruction&ScreeningKit(#K1617-1).ItcontainsaHIS3nutritionalreportergene,locateddownstreamofamultiplecloningsite(MCS)andtheminimalpromoteroftheHIS3locus(PminHIS3).Cis-actingDNAsequences,orDNAtargetelements,canbeinsertedintotheMCSandusedasbaitstoscreenGAL4AD/cDNAfusionlibrariesforproteinsthatinteractwiththetargetsequence.Aprotein-DNA(orone-hybrid)interactioncanbedetectedbyperformingtheassayinayeaststrainsuchasY187thatisauxotrophicforhistidine.Positiveone-hybridinteractionsdriveexpressionoftheHIS3reportergene,whichenablesthehostcelltogrowonhistidine-deficientmedia.Intheabsenceofactivation,theconstitutiveHIS3expressionfromPminHIS3isverylow.Duringlibraryscreening,theleakyexpressionofHIS3iscontrolledbyadding3-amino-1,2,4-triazole(3-AT)tothemedium.Theconcentrationof3-ATneededtofullysuppressleakyHIS3expressionmustbedeterminedempiricallyforeachDNAtargetelement.pHIS2canbemaintainedinbothyeastandbacteria.Itcontainsanautonomousreplicationsequence(ARS4)andTRP1nutritionalmarkerforreplicationandselectioninyeast(1,2);itcontainsaColE1originandakanamycinresistancegene(Kanr)forpropagationandselectioninE.coli.ThecentromericsequenceCEN6ensurespropersegregationoftheplasmidduringcelldivisioninyeast(1,2).UseTousepHIS2inaone-hybridassay,cloneoneormorecopiesofacis-actingDNAtargetsequenceintotheMCS.ThenintroducetheplasmidintocompetentyeastcellsusingthetransformationprotocolintheBDMatchmakerLibraryConstruction&ScreeningKitsUserManual(PT3529-1).IncontrasttotheoriginalBDMatchmakerOne-HybridSystem,thisreportervectordoesnotneedtobeintegratedintotheyeastgenome.Instead,itismaintainedasanepisomethroughouttheassay.InsertingyourtargetelementmayalterthelevelofbackgroundHIS3expression.Therefore,constructs首ldbetestedforbackground(leaky)HIS3expressionbeforeyoustartaone-hybridanalysis.BackgroundgrowthduetoleakyHIS3expressioniscontrolledbyadding3-ATtotheselectionmedium,asdescribedintheUserManual(PT3529-1).LocationofFeaturesMultiplecloningsites:141HIS3gene:152811FragmentcontainingtheHIS33'UTR&Terminationsequence:8121446TRP1gene:28553529FragmentcontainingtheColE1E.colioriginofreplication:41654615Kanamycin-resistancegene:56054811CEN6/ARS4sequences:62545737

參與評論

全部評論(0條)

更多資料

該會員其他資料
D12492說明書
CYGNUS F050說明書
HIV P24 ELISA KIT 說明書
research diets D12451說明書
research diets d12450b說明書
omicronbio產(chǎn)品
Smartox-biotech產(chǎn)品
phytotechlab產(chǎn)品
美天旎macs代理
polysciences產(chǎn)品目錄

在線留言

換一張?
取消