Clontech AH109说明书

Clontech AH109说明书

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ClontechAH109说明书，AH109说明书，ClontechAH109AH109型号载体名称出品公司载体用途VNC0491AH109Clontech配套酿酒酵母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.

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Clontech AH109说明书

ClontechAH109说明书，AH109说明书，ClontechAH109AH109型号载体名称出品公司载体用途VNC0491AH109Clontech配套酿酒酵母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.[详细]
2018-09-29 10:03
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Clontech pTK-hyg （VSC0467）说明书

pTK-hyg型号载体名称出品公司载体用途VSC0467pTK-hygClontech四环素调控系统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质粒图谱:[详细]
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Clontech酵母双杂交系统 pCL1说明书

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Clontech 酵母双杂交系统 pGBKT7说明书

Clontech酵母双杂交系统pGBKT7，酵母双杂交系统pGBKT7，ClontechpGBKT7pGBKT7产品参数:MammalianSelection：TRP1（HIS3selection）载体抗性:卡那霉素（Kanamycin）说明书:质粒图谱:[详细]
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酵母单杂交系统ClontechpHis2说明书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[详细]
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说明书

说明书[详细]
2007-05-30 00:00
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Clontech AH109说明书

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