学科分类
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29 个结果
  • 简介:Changesinthedistributionof1P1-antigeninthedevelopingchickretinahavebeenexaminedbyindriectimmunofluorescencestainingtechniqueusingthenovelmonoclonalantibody(MAb)1P1.Expressionofthe1P1antigenwasfoundtoberegulatedinradialaswellasintangentialdimensionoftheretina,beingpreferentiallyorexclusivelylocatedintheinnerandouterplexiformlayersoftheneuralretinadependingonthestagesofdevelopment,Withtheonsetoftheformationoftheinnerplexiformlayer1P1antigenbecomesexpressedintheretina.Withprogressingdifferentiationoftheinnerplexiformlayer1P1immunofluorescencerevealed2subbandsatE9and6subandsatE18,Atpostnatalstages(afterP3)immunoreactivitywasreducedinaninside-outsidesequenceleadingtothecompleteabsenceofthe1P1antigeninadulthood.1P1antigenexpressionintheouterplexiformlayerwasalsosubjecttodevelopmentalregulation.Thespation-temporalpatternof1P1antigenexpressionwascorrelatedwiththetimecourseofhistologicaldifferentationofchickretina,namelythesynapserichplexiformlayers.Whetherthe1P1antigenwasfunctionallyinvolvedindendriteextensionandsynapseformationwasdiscussed.

  • 标签: 鸡胚 视网膜发育 网织层 IP1抗原 表达 时空分布
  • 简介:Arabidopsisthaliana嘘deacetylase1(AtHD1或AtHDA19),酵母RPD3个相当或相同事物,在植物许多生理、发展过程个全球管理者。尽管有为在植物基因规定和开发AtHD1个角色基因证据,AtHD1生物化学、细胞性质糟糕被理解。这里,我们在vivo报导AtHD1细胞本地化模式并且嘘在vitro项deacetylase活动。绿荧光灯蛋白质(GFP)短暂、稳定表示在洋葱房间标注了AtHD1并且分别地,在转基因Arabidopsis根,种子和叶子表明AtHD1在euchromatic区域大概在原子核局部性并且从核排除。AtHD1本地化模式与涉及核形成和transgenessilencing并且分别地重复了DNA元素AtHD2和AtHDA6那些不同。另外,histdeacetylase活动试金证明在细菌生产recombinantAtHD1示威了特定嘘在vitro项deacetylase活动。数据建议AtHD1原子蛋白质并且拥有嘘为对植物生长和开发重要全球transcriptional规定负责项deacetylase活动。

  • 标签: 组蛋白脱乙酰基1 常染色体 基因抑制 植物
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  • 简介:TheyeastHAL1genewasintroducedintoArabidopsisthalianabyAgrobacteriumtumefaciens-mediatedtransformationwithvacuuminfiltrationunderthecontrolofCaMV35Spromoter.Thirty-threeindividualkanamycinresistantplantswereobtainedfrom75,000seeds.SouthernblottinganalysisindicatedthatHAL1genehadbeenintegratedintoallofthetransgenicplants'genomes.ThecopynumberofHAL1geneintransgenicplantswasmostly1to3bySouthernanalysis.Phenotypesoftransgenicplantshavenodifferenceswithwildtypeplants.Severalsamplesoftransformantswereself-pollinated,andprogeniesfromtransformedandnon-transformedplants(controls)wereevaluatedforsalttoleranceandgeneexpression.MeasurementofconcentrationsofintracellularK+andNa+showedthattransgeniclineswereabletoretainlessNa+thanthatofthecontrolundersaltstress.ResultsfromdifferenttestsindicatedtheexpressionofHAL1genepromotesahigherlevelofsalttoleranceinvivointhetransgenicArabidopsisplants.

  • 标签: 耐盐碱性 HAL1基因 基因工程
  • 简介:Galα(1,3)Gal(galepitope)isacarbohydrateepitopeandsynthesizedinlargeamountbyα(1,3)galactosyltransferase[α(1,3)GT]enzymeonthecellsoflowermammaliananimalssuchaspigsandmice.Humanhasnogalepitopeduetotheinactivationofα(1,3)GTgenebutproducesalargeamountofantibodies(anti-Gal)whichrecognizeGalα(1,3)Galstructuresspecifically.Inthisstudy,areplicationdeficientrecombinantadenoviralvectorAd5sGTcontainingpigα(1,3)GTcDNAwasconstructedandcharacterized.Adenoviralvector-mediatedtransferofpigα(1,3)GTgeneintohumantumorcellssuchasmalignantmelanomaA375,stomachcancerSGC-7901,andlungcancerSPC-A-1wasreportedforthefirsttime.ResultsshowedthatGalepitopedidnotincreasethesensitivityofhumantumorcellstohumancomplement-mediatedlysis,althoughhumancomplementactivationandthebindingofhumanIgGandIgMnaturalantibodiestohumantumorcellswereenhancedsignificantlyafterAd5sGTtransduction.AppearanceofgalepitopeonthehumantumorcellschangedtheexpressionofcellsurfacecarbohydratesreactingwithUlexeuropaeusI(UEAI)lectins,Viciavillosaagglutinin(VVA),Arachishypogaeaagglutinin(PNA),andGlycinemaxagglutinin(SBA)todifferentdegrees.Inaddition,noeffectofgalepitopeonthegrowthinvitroofhumantumorcellswasobservedinMTTassay.

  • 标签: 腺病毒载体 半乳糖转移酶 GAL α(1 3) Gao 基因表达
  • 简介:SomerecentstudiesindicatedthatGABAergicsystemisinvolvedinmammalianspermacrosomereaction(AR),butdirestevidencepertainingtotheexpressionofgatlinmammalianspermisnotyetdemonstrated.Inthisstudy,weevaluatedthepresenceof67kDaGAT1proteinandmRNAinrattestisbyWesternblottingandreversetranscription-polymerasechainreaction.Meanwhile,immunohistochemicalandimmunofluorescentanalysesalsoidentifiedGAT1proteinontheelongatedspermatidandsperm.Theseresultsindicatedthatrattestisisanovelsiteofgatlexpression.FurtherstudiesshouldbetakentoexploretheroleofGAT1proteinonspermacrosomereaction.

  • 标签: 大鼠 精子 睾丸 GAT1 表达 GABA
  • 简介:以便学习TGF-beta1结构功能细节,重组体老鼠TGF-beta1成熟形式在细菌被表示。TGF-beta1(氨基酸279-390)112氨基酸carboxyl终端部分合成被个可诱导基因表示系统基于抗菌素T7RNA控制。这个系统允许重组体TGF-beta1活跃、选择合成。在减少条件下面在SDS-polyacrylamide胶化上决定表示TGF-alpha1单体分子量大约13kD。连续净化洗与纯化步足够净化表示产品同质单个胶化过滤结合了。氨基终端定序表明重组体蛋白质N终端与出版数据相同。在西方污点分析,重组体多肽对polyclonalTGF-beta1抗体显示出优秀antigenicity。在这研究表示成熟重组体老鼠TGF-beta1提供个有用工具因为未来详细说明了结构、功能研究。

  • 标签: 转化生长因子-β1 重组表达 包涵体 提纯 抗原性
  • 简介:Thereareabout17chromosomesinyeastSaccharomycescerevisiae.Amiddlesizedchromosome,chromosomeV,waschoseninthisworkforstudyingandconstructingthephysi-calmaps.ChromosomeVfromstrainA364awasisolatedbypulsed-fieldgradientgelelectrophoresis(PFGE).GelslicescontainingchromosomeVDNAweredigestedwithtworarecuttingenzymes,NotⅠandSfiⅠ,andthree6-Ntrecognizingenzymes,SmaⅠ,SstⅡandApaⅠ.Severalstrategies-partialorcompletedigestions,digestionwithdifferentsetsoftwoenzymes,andhybrid-izationwithclonedgeneticallymappedprobes(CAN1,URA3,CEN5,PRO3,CHO1,SUP19,RAD51,RAD3)——wereusedtoaligntherestrictionfragments.Thereare9,9,15,17,and20sitesforNotⅠ,SfiⅠ,SmaⅠ,SstⅡandApaⅠrespectivelyinthemapoftheA364achromosomeV.Itstotallengthwascalculatedtobe620Kb(Kilo-bases).Thedistributionsofthecuttingsitesforthesefiveenzymesthroughthewholechromosomearenotuniform.Acomp-arisonbetweenthephysicalmapandthegeneticmapwasalsomade.

  • 标签: YEAST chromcsome V macro-physical map DNA
  • 简介:尽管许多进步在理解花机关身份怎么在花发展期间坚定被取得了,更少花机关怎么在晚花发展阶段被详细描述被知道。这里,我们描述新奇花异种,起皱花瓣和stamens1(wps1),它在花瓣和雄蕊发展显示出缺点。基因分析显示那wps1异种相应于在染色体3长手臂个单个后退地点。在wps1异种机关发展在野类型类似于那,并且异种malfunction在晚发展阶段开始,显示花瓣和雄蕊外观上个缺点。在成熟花,在异种花瓣和雄蕊细丝被起皱或合拢,并且在花瓣和雄蕊细丝L1层下面的细胞形态学反常。花机关身份基因表达式模式没与野类型相比在wps1异种被影响,这被发现,与所有花机关未改变开发致。而且,在花瓣不同类型表皮房间身份被维持。组织学分析证明在wps1花,所有花瓣不规则地被合拢,并且在花瓣有系结构,当内部房间形状和安排畸形、未组织起来时。把结果基于这些,我们建议Wps1下游地行动到班B花机关身份基因,并且工作在迟了花期间调制细胞区别发展阶段。

  • 标签: 花瓣 雄蕊 莲花 植物
  • 简介:GABAtransporter1(GAT1)takesimportantrolesinmultiplephysiologicalprocessesthroughtheuptakeandreleaseofGABA,buttheregulationofGAT1geneexpressionindifferenttissuesisrarelyknown.Toaddressthequestion,first,5'RapidamplificationofcDNAend(RACE)wasusedtodetermineGAT1transcriptionalstartingsitesinneonatalmousecerebralcortexandintestine,adultmousebrainandadultrattestis.Theproductsof5'RACEwereconfirmedbyDNAsequencing.WefoundthatthetranscriptofGAT1inneonatalmousecerebralcortexandadultmousebrainstartsatthesamesite(insideofexon1),whileinmouseintestine,GAT1startstranscriptioninintron1,andinrattestis,thetranscriptofGAT1hasanadditionaluntranslationexontothe5'direction.

  • 标签: GABA转移子1 复制开始位点 5'RACE 组织特异性
  • 简介:P28,a28kDproteinfromtoad(Bufobufogargarizans)oocytes,wasidentifiedbyusingP13^suc1-agaroseaffinitychromatography.Sequencehomologyanalysisofthefull-lengthcDNAofP28(GeneBankaccessionnumber:AF314091)indicatedthatitencodesaproteincontaining224amino-acidswithabout55%iden-titiesandmorethan70%positivestoencodesaproteincontaining224amino-acidswithabout55%iden-titiesandmorethan70%positivestohuman,ratormouseUCH-L1,andcontainshomologicalfunctionaldomainsofUCHfamily.Anti-p28monoclonalantibody,oninjectingintotheoocytes,couldinhibittheprogesterone-inducedresumptionofmeioticdivisioninadose-dependentmanner.TherecombinantproteinP28showedsimilarSDS/PAGEbehaviorstothenativeone,andpromotedubiquitinethylesterhydrolysis,aclassicalcatalyticreactionforubiquitincarboxylterminalhydrolases(UCHs).Theresultsinthispaperrevealthatanovelprotein,p28,existsinthetoadoocytes,isaUCHLlhomolog,wasengagedintheprocessofprogesterone-inducedoocytematurationpossiblythroughaninvolvementinproteinturnoveranddegradation.

  • 标签: 蟾蜍 卵母细胞成熟 p28 cDNA克隆 重组表达 蛙卵
  • 简介:γ-AminobutyricacidandGABAergicreceptorswerepreviouslyreportedtobedistributedinreproductivesystemsbesidesCNSandpredictedtoparticipateinthemodulationoftesticularfunction.γ-Aminobutyricacidtransporterwasimplicatedtobeinvolvedinthisprocess.However,thepotentialroleofγ-aminobutyrictransporterintestishasnotbeenexplored.Inthisstudy,weinvestigatedtheexistenceofmouseγ-aminobutyricacidtransportersubtypeI(mGAT1)intestis.Wild-typeandtransgenicmice,whichoverexpressingmGAT1inavarietyoftissues,especiallyintestis,wereprimarilystudiedtoapproachtheprofileofmGAT1intestis.MicewithoverexpressedmGAT1developnormallybutwithreducedmassandsizeoftestisascomparedwithwild-type.Testicularmorphologyoftransgenicmiceexhibitedovertabnormalitiesincludingfocaldamageofthespermatogenicepitheliumaccompaniedbycapillariesproliferationandincreaseddiameterofseminiferoustubuleslumen.Reducednumberofspermatidswasalsofoundinsomeseminiferoustubules.OurresultsclearlydemonstratethepresenceofGAT1inmousetestisandimplythatGAT1ispossiblyinvolvedintesticularfunction.

  • 标签: 睾丸形态 GAT1 过表达 GABA γ-氨基丁酸转运蛋白
  • 简介:ASK1(ARABIDOPSIS象SKP1样)蛋白质招募ubiquitinligase建筑群指向SCF(Skp1-Cullin-F盒子蛋白质)个批评部件为由26Sproteosome降级蛋白质。为了调查蛋白质,那被调停ASK1解朊作用小径在Arabidopsis花影响,我们用二维电气泳动(2-DE)比较了Arabidopsis野类型和ask1异种花芽proteomes。有在与野类型花相比ask1异种花更高或更低丰富十个蛋白质点被切除并且使分析遭到了步集体spectrometry(MS)。结果证明他们涉及相片形态发生,生理节奏摆动,翻译以后过程,压力反应和房间扩大或延伸蛋白质,建议那些过程在ask1异种被影响。基因也是的这些抄本层次基于Affymetrix基因芯片比较了微数组数据。没有重要差别为大多数基因被观察,建议有在ask1异种累积提高层次蛋白质能条调停ASK1解朊作用小径调整候选人目标。阐明这些结果帮助在Arabidopsis发展过程并且也ASK1多种功能表明关于基因功能学习蛋白质层次重要性和必要性。

  • 标签: 蛋白质 水解途径 电位 生物研究
  • 简介:Activehost-pathogeninteractionstakeplaceduringinfectionofhumanimmunodeficiencyvirustype1(HIV-1).Outcomesoftheseinteractionsdeterminetheefficiencyofviralinfectionandsubsequentdiseaseprogression.HIV-infectedcellsrespondtoviralinvasionwithvariousdefensivestrategiessuchasinnate,cellularandhumoralimmuneantiviralmechanisms.Ontheotherhand,thevirushasalsodevelopedvariousoffensivetacticstosuppressthesehostcellularresponses.Amongmanyoftheviraloffensivestrategies,HIV-1viralauxiliaryproteins(Tat,Rev,Nef,Vif,VprandVpu)playimportantrolesinthehost-pathogeninteractionandthushavesignificantimpactsontheoutcomeofHIVinfection.OneofthebestexamplesistheinteractionofVifwithahostcytidinedeaminaseAPOBEC3G.AlthoughspecificrolesofotherauxiliaryproteinsarenotaswelldescribedasVif-APOBEC3Ginteraction,itisthegoalofthisbriefreviewtosummarizesomeofthepreliminaryfindingswiththehopetostimulatefurtherdiscussionandinvestigationinthisexhilaratingareaofresearch.

  • 标签: 爱滋病病毒-1 辅助蛋白 病毒感染 中国 病原体
  • 简介:ToexplorethemolecularmechanismofchromatinremodelinginvolvedintheregulationoftranscriptionalactivationofspecificgenesbyamyogenicregulatoryfactorMyogenin,weusedNIH3T3fibroblastswithastablyintegratedH1.1-GFPfusionproteintomonitorhistoneH1movementdirectlybyfluorescencerecov-eryafterphotobleaching(FRAP)inlivingcells.TheobservationfromFRAPexperimentswithmyogenintransfectedfibroblastsshowedthattheexchangerateofhistoneH1inchromatinwasobviouslyincreased,indicatingthatforcedexpressionofexogenousMyogenincaninducechromatinremodeling.Thehyper-acetylationofhistonesH3andH4frommyogenintransfectedfibroblastswasdetectedbytriton-acid-urea(TAU)/SDS(2-D)electrophoresisandWesternblotwithspecificantibodiesagainstacetylatedN-terminiofhistonesH3andH4.RT-PCRanalysisindicatedthatthenAChRa-subunitgenewasexpressedinthetrans-fectedfibroblasts.TheseresultssuggestthattheexpressionofexogenousMyogenincaninducechromatinremodelingandactivatethetranscriotionofMvogenin-targetedgeneinnon-musclecells.

  • 标签: 染色体 组蛋白 染色体改变 乙酰化
  • 简介:Inmacrophages,theaccumulationofcholesterylesterssynthesizedbytheactivatedacyl-coenzymeA:cholesterolacyltransferase-1(ACAT1)resultsinthefoamcellformation,ahallmarkofearlyatheroscleroticlesions.Inthisstudy,withthetreatmentofaglucocorticoidhormonedexamethasone(Dex),lipidstainingresultsclearlyshowedthelargeaccumulationoflipiddropletscontainingcholesterylestersinTHP-1-derivedmacrophagesexposedtolowerconcentrationoftheoxidizedlow-densitylipoprotein(ox-LDL).Morenotably,whentreatedtogetherwithspecificanti-ACATinhibitors,theabundantcholesterylesteraccumulationwasmarkedlydiminishedinTHP-l-derivedmacrophages,confirmingthatACATisthekeyenzymeresponsibleforintracellularcholesterylestersynthesis.RT-PCRandWesternblotresultsindicatedthatDexcausedup-regulationofhumanACAT1expressionatboththemRNAandproteinlevelsinTHP-1andTHP-1-derivedmacrophages.TheluciferaseactivityassaydemonstratedthatDexcouldenhancetheactivityofhumanACAT1geneP1promoter,amajorfactorleadingtotheACAT1activation,inacell-specificmanner.Furtherexperimentalevidencesshowedthataglucocorticoidresponseelement(GRE)locatedwithinhumanACAT1geneP1promotertoresponsetotheelevationofhumanACAT1geneexpressionbyDexcouldbefunctionallyboundwithglucocorticoidreceptor(GR)proteins.ThesedatasupportedthehypothesisthattheclinicaltreatmentwithDex,whichincreasedtheincidenceofatherosclerosis,mayinpartduetoenhancingtheACAT1expressiontopromotetheaccumulationofcholesterylestersduringthemacrophage-derivedfoamcellformation,anearlystageofatherosclerosis.

  • 标签: 巨噬细胞 地塞米松 泡沫细胞 人类 基因 表达
  • 简介:WereportedinthismanuscriptthatTGF-β1inducesapoptosisinAML12murinehepatocytes,whichisassociatedwiththeactivationofp38MAPKsignalingpathway.SB202190,aspecificinhibitorofp38MAPK,stronglyinhibitedtheTGF-β1-inducedapoptosisandPAI-1promoteractivity.TreatmentofcellswithTGF-β1activatesp38.Furthermore,over-expressionofdominantnegativemutantp38alsoreducedtheTGF-β1-inducedapoptosis.Thedataindicatethattheactivationofp38isinvolvedinTGF-β1-mediatedgeneexpressionandapoptosis.

  • 标签: 转化生长因子Β 细胞凋亡 P38 肝细胞 信号传导
  • 简介:MembersofBcl-2familyofproteinsareregulatorsofcelldeaththatcanbegroupedintosubfamiliesofprosurvivalandproapoptoticmolecules.Theyarecharacterizedbythepresenceofseveralconservedmotifs,knownastheBcl-2homology(BH)domains,designatedBH1,BH2,BH3andBH4.MutagenesisandstructuralstudiesrevealedthattheBHdomainsareimportantfunctionaldomainsthatarealsorequiredfordimerizationfunction.Recently,asubfamilyofproapoptoticmoleculesonlycontainsBH3motifhasbeenidentifiedsuggestingBH3domainalonemaybesufficientformediatingproapoptoticfunctionamong

  • 标签: 前凋亡蛋白 map-1 多bcl-2同源域 bax相关蛋白 特性
  • 简介:TRAF2isacriticaladaptormoleculeforTNFreceptorsininflammatoryandimmunesignaling.Uponreceptorengagement,TRAF2isrecruitedtoCD40andtranslocatestolipidraftsinaRINGfinger-dependentprocess,whichenablestheactivationofdownstreamkinases.TRAF1candisplaceTRAF2andCD40fromraftfractions,anditpromotestheabilityofTRAF2tosustainsignalactivation.ReplacementoftheRINGfingerofTRAF2witharaft-targetingsignalrestoresJNKactivationandassociationwiththecytoskeletalproteinFilamin,butnotNF-KBactivation.TRAF1-/-dendriticcellsshowattenuatedresponses

  • 标签: TRAF2细胞内定位 TRAF1调节 信号转导机制
  • 简介:Ionizingradiationisoneofthemosteffectivetoolsincancertherapy.Inapreviousstudy,wereportedthatproteintyrosinekinase(PTK)inhibitorsmodulatetheradiationresponsesinthehumanchronicmyelogenousleukemia(CML)celllineK562.Thereceptortyrosinekinaseinhibitor,genistein,delayedradiation-inducedcelldeath,whilenon-receptertyrosinekinaseinhibitor,herbimycinA(HMA)enhancesradiation-inducedapoptosis.Inthisstudy,wefocusedonthemodulationofradiation-inducedcelldeathbygenisteinandperformedPCR-selectsuppressionsubtractivehybridization(SSH)tounderstanditsmolecularmechanism.Weidentifiedhumanthymidinekinase1(TK1),whichiscellcycleregulatorygeneandconfirmedexpressionofTK1mRNAbyNorthernblotanalysis.ExpressionofTK1mRNAandTK1enzymaticactivitywereparallelintheirincreaseanddecrease.TK1isinvolvedinG1-Sphasetransitionofcellcycleprogression.Incellcycleanalysis,weshowedthatradiationinducedG2arrestinK562cellsbutitwasnotabletosustain.However,theadditionofgenisteintoirradiatedcellssustainedaprolongedG2arrestupto120h.Inaddition,theexpressionofcellcycle-relatedproteins,cyclinAandcyclinB1,providedtheevidencesofG1/SprogressionandG2-arrest,andtheirrelationshipwithTK1incellstreatedwithradiationandgenistein.TheseresultssuggestthattheactivationofTK1maybecriticaltomodulatetheradiation-inducedcelldeathandcellcycleprogressioninirradiatedK562cells.

  • 标签: 染料木素 胸腺嘧啶核苷激酶1 活化 辐射 细胞凋亡 K562