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• Clontech AH109說(shuō)明書(shū)

  ClontechAH109說(shuō)明書(shū)，AH109說(shuō)明書(shū)，ClontechAH109AH109型號(hào)載體名稱出品公司載體用途VNC0491AH109Clontech配套釀酒酵母hissectionprovidesdetailedphenotypesoftheyeaststrainsincludedwithMATCHMAKERSystem3andMATCHMAKERGAL4cDNAandGenomicLibraries.Foradditionalinformationonthegrowthandmaintenanceofyeast,seetheYPH,ChapterIII.WealsorecommendGuthrie&Fink’sGuidetoYeastGeneticsandMolecularBiology（1991）andHeslot&Gailardin’sMolecularBiologyandGeneticEngineeringofYeasts（1992）.A.YeastHostStrainsThecompletegenotypesofAH109,Y187,andCG-1945areprovidedinTableII.Allstrainsaregal4andgal80;thispreventsinterferenceofnativeregulatoryproteinswiththeregulatoryelementsinthetwo-hybridsystem.1.UseAH109asthehoststrainifyouplantoscreenanAD/libraryusingHIS3,ADE2,andMEL1.2.System3UsersOnly:UseY187asthehoststrainifyouplantotestforaninteractionbetweentwoknownproteinsusingthelacZreporteronly.Inaddition,useY187asamatingpartnertoverifyproteininteractions.3.LibraryUsersOnly:UseCG-1945asthehoststrainifyouplantoseparateDNA/baitandAD/libraryplasmidsbycycloheximidecounterselection.Alternatively,usepGBKT7toconstructyourbait.ThisDNA-BDvectorcontainsakanamycinresistancemarker;therefore,vectorscanbeseparatedinE.coliwithoutcycloheximidecounterselection.[詳細(xì)]

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  產(chǎn)品樣冊(cè)

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• Clontech pTK-hyg （VSC0467）說(shuō)明書(shū)

  pTK-hyg型號(hào)載體名稱出品公司載體用途VSC0467pTK-hygClontech四環(huán)素調(diào)控系統(tǒng)DescriptionpTK-Hygisaselectionvectorthatconfershygromycinresistanceinmammaliancellsfortheselectionofstablytransformedcells.pTK-Hygisespeciallyusefulforselectionofdouble-stablecelllinesusingtheTet-OnorTet-OffGeneExpressionSystemsbecausetheabsenceofanenhancerelementonpTK-HygpreventstheunwantedactivationofpTRE-andpBI-derivedplasmidsuponcointegrationintothehostcell'sgenome.UsepTK-Hygcanbecotransfectedintothehostcelllinewithanexpressionplasmidthatcontainsageneofinterestusinganystandardtransfectiontechnique.Mostmammaliancelllinesrequire200g/mlofHygromycintoselectforstablytransformedcells.LocationofFeaturesHSVTKpolyAsignals:14731478&14861491（complementary）Hygr（hygromycinresistancegene）:15372574（complementary）PHSVTK（HSVTKpromoter）:25882835（complementary）pUCoriginofreplication:3152379mpr（ampicillinresistancegene;beta-lactamase）:39434803PropagationinE.coliSuitablehoststrains:DH5aandothergeneralpurposestrains.Selectablemarker:plasmidconfersresistancetoampicillin（50g/ml）onE.colihosts.E.colireplicationorigin:pUC質(zhì)粒圖譜:[詳細(xì)]

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• Clontech酵母雙雜交系統(tǒng) pCL1說(shuō)明書(shū)

  MATCHMAKERGAL4Two-HybridVectorsHandbookpCL1型號(hào)載體名稱出品公司載體用途VJC0484pCL1Clontech酵母雙雜交系統(tǒng)產(chǎn)品參數(shù):測(cè)序引物：CMV-F載體抗性:氨芐青霉素（Ampicillin）載體描述:pCL1isapositivecontrolplasmidthatencodesthefulllength,wild-typeGAL4protein.TheGAL4proteinactivatesreportergenesunderthecontrolofaGAL4-responsiveelement（UASG）.Thus,pCL1isusedasapositivecontrolforthetranscriptionassayinGAL4-basedMATCHMAKERTwo-HybridSystems.ThisvectorisaderivativeofYCp50referencedinFields&Song,1989.pCL1hasnotbeensequenced.質(zhì)粒圖譜:[詳細(xì)]

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• Clontech 酵母雙雜交系統(tǒng) pGBKT7說(shuō)明書(shū)

  Clontech酵母雙雜交系統(tǒng)pGBKT7，酵母雙雜交系統(tǒng)pGBKT7，ClontechpGBKT7pGBKT7產(chǎn)品參數(shù):MammalianSelection：TRP1（HIS3selection）載體抗性:卡那霉素（Kanamycin）說(shuō)明書(shū):質(zhì)粒圖譜:[詳細(xì)]

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• Clontech pBridge酵母三雜交系統(tǒng)說(shuō)明書(shū)

  ClontechpBridge酵母三雜交系統(tǒng)說(shuō)明書(shū)載體描述:pBridgeexpressestwoproteins:aDNA-bindingdomainfusion,andanadditionalprotein.pBridgethusallowsestablishmentofthree-hybridsystemswhenusedincombinationwithanactivationdomainfusionvectorandyeaststrainsfromanyofClontech'sGAL4-basedtwo-hybridsystems,includingMatchmakerGold.ThisvectorgeneratesahybridproteinthatcontainsthesequencesfortheGAL4DNA-bindingdomain（DNA-BD）andthesequenceclonedintoMCSI.ThefusionproteinisexpressedinyeasthostcellsfromtheconstitutiveADH1promoter;transcriptionisterminatedattheADH1transcriptionterminationsignal.Thehybridproteinistargetedtotheyeastnucleusbynuclearlocalizationsequences（NLS）thatareanintrinsicpartoftheGAL4DNA-BD.AnadditionalgeneofinterestcanbeclonedintoMCSIIwhichislocateddownstreamofanHAepitopeandasecondNLS.TheresultingfusionproteinisconditionallyexpressedfromtheMet25promoterinresponsetomethioninelevelsinthemedium;i.e.,itisrepressedinthepresenceof1mMmethionineandexpressedintheabsenceofmethionine.質(zhì)粒圖譜:[詳細(xì)]

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• 酵母單雜交系統(tǒng)Clontech pHis2說(shuō)明書(shū)

  酵母單雜交系統(tǒng)ClontechpHis2說(shuō)明書(shū)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[詳細(xì)]

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• 說(shuō)明書(shū)

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  選購(gòu)指南

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