简介:WedevelopedanRfunctionnamed'microarrayoutlierfilter'(MOF)toassistintheidentificationoffailedarrays.Insortingagroupofsimilararraysbythelike-lihoodoffailure,twostatisticalindiceswereemployed:thecorrelationcoefficientandthepercentageofoutlierspots.MOFcanbeusedtomonitorthequalityofmi-croarraydataforbothtroubleshooting,andtoeliminatebaddatasetsfromdown-streamanalysis.Thefunctionisfreelyavaliableathttp://www.wriwindber.org/applications/mof/.
简介:MYB蛋白质在真核细胞的有机体起重要作用。在植物,R1R2R3类型MYB蛋白质在房间周期控制工作。然而,R2R3类型MYB蛋白质是否也涉及房间部门过程,仍然保持未知。这里,我们报导那R2R3类型抄写因素基因,AtMYB59,涉及房间周期前进和根生长的规定。AtMYB59蛋白质在洋葱的原子核是局部性的表皮的房间并且transactivation活动。在酵母房间的AtMYB59的表示压制房间增长,和transformants与更长的房间有更多的原子核和更高的aneuploidDNA内容。在AtMYB59的保存领域的变化在酵母细胞生长上废除它的效果。在同步Arabidopsis房间暂停,AtMYB59基因明确地在房间周期前进期间在S阶段被表示。表示和promoter-GUS分析表明AtMYB59基因富有地在根被表示。转基因的植物overexpressingAtMYB59更短的根与野类型的植物(Arabidopsis就职Col-0)相比,并且在在根尖端的有丝分裂的房间的一半附近在中期。相反地,空变异的myb59-1比关口在中期让更长的根和更少有丝分裂的房间,建议那AtMYB59可以由扩大有丝分裂的房间的中期禁止根生长。AtMYB59调整许多下游的基因,包括CYCB1;1基因,可能通过到MYB应答的元素的绑定。这些结果在细胞周期规定和植物根生长为AtMYB59支持一个角色。
简介:死亡联系域的蛋白质Daxx施加包括调停的许多报导功能经由激活c6月N终端从FasL发信号到apoptosisapoptosis的kinase(JNK),正式就职和抑制,和染色质改变的规定。它原来从酵母被克隆用船边交货对相似物体之连续感觉而形成心像口的细胞内部的尾巴的二混血儿的屏幕诱饵。而许多起始的报告仍然保持争论,Daxx在一系列压力信号包括UVirradiation,过氧化氢治疗和TGF-β治疗触发的apoptosis的规定起重要作用,是清楚的。在这评论,我们在Axinbeing上集中连接Daxx到p53的一个拴住的因素。
简介:CCAAT/enhancer有约束力的蛋白质α(C/EBPα)是transcriptional禁止细胞增殖的规章的因素,和其他的翻译开始生产二多肽,C/EBPαp30并且C/EBPαp42。由表示介绍,C/EBPαp30specifically禁止了基因的一个唯一的集合,这被揭示,包括MPP11,p84N5和SMYD2,它没被C/EBPαp影响42在两根QSG-7701hepatocyte房间线和QGY-7703hepatoma,cells.Semi量的RT-PCR分析独立地证实了这些结果。Chromatinimmunoprecipitation试金显示出30比C/EBPαp更强烈绑了在这些基因的倡导者的那C/EBPαp42。在C/EBPα是调整的down的临床的hepatoma在样品,所有三基因明确地由30在上面的C/EBPαp禁止了调整。然而,MPP11,p84N5和SMYD2genes的压抑可能直接不涉及C/EBPαp30-mediated生长抑制。我们的数据建议30调整的那C/EBPαp目标基因的一个唯一的集合并且多于C/EBPαp的dominant-negativeregulator42。
简介:应用DNA聚合酶链式反应(PCR)技术,对p16抑癌基因(CDKN2)进行体外定点突变,在p16cDNA中引入第48位密码子CCG(Pro)→CTG(Leu)和第74位密码子GAC(Asp)→AAC(Asn)突变,构建了p16-P48L和p16-D74N突变体。野生型和突变型p16cDNA克隆于pcDNA3构建pCMV-p16、pCMV-p16P48L和pCMV-p16D74N真核表达载体,导入纯合缺失p16基因的人肺癌细胞株H460,经RNA点杂交、RNA印迹和细胞免疫化学染色,检测到P16表达。通过比较表达野生型和突变型P16的H460细胞在~3H-TdR掺入及细胞所在周期的差异,证实P16表达抑制细胞进入S期,而P48L和D74N突变体对细胞进入S期没有什么影响,提示P48L和D74N突变导致P16蛋白功能丧失。
简介:在Saccharomycescerevisiae,必要基因CDC13编码telomeric遗传上并且身体上与Stn1p和Ten1p交往的搁浅单人赛的DNA有约束力的蛋白质,并且为telomere结束保护和telomere长度控制被要求。Ten1由参予telomere长度规定和染色体结束保护的分子的机制留下逃犯。在这个工作,我们用净化的recombinantCdc13p和Ten1p在胶化过滤分析观察了Cdc13p和Ten1p的一个弱相互作用。Ten1p本身展出一项弱DNA有约束力的活动,但是提高telomericTG1鈥吗?Cdc13p的DNA有约束力的能力。Cdc13p是有Ten1p的co-immunoprecipitated。在变异的ten1-55或ten1-66房间,在Ten1p和Cdc13p之间的损害相互作用与telomericDNA导致长得多的telomeres,以及Cdc13p的一个减少的协会。一致地,Ten1-55和Ten1-66异种蛋白质没能刺激telomeric在vitro的Cdc13p的DNA有约束力的活动。这些结果建议Ten1p提高telomericCdc13p到的DNA有约束力的活动否定地调整telomere长度。
简介:TheundecapeptidesubstanceP(SP)wasshowntobeintimatelyinvolvedinboththestructuralandfunctionalaspectsoftheanteriorpituitary.Yet,inadditiontoitsinfluencesonhormonalsecretion,SPmaywellpossessmoreactionsinthismastergland.ThepresentstudywasfthereforeaimedtoinvestigatetheeffectofSPontheproliferationofratanteriorpituitarycellsinprimaryculture,ItwasfoundthatSPcoulddose-dependentlyincreasetheincorporationoftritiatedthymidine(3H-TdR)intoculturedanteriorpituitarycells.OthermammaliantachykininssuchasneurokininAandneurokininBhadsimilareffectbuttovaryingdegrees.TheequipotentanalogueofSP,Norleucine^11-SP(Nle^11-SP),alsoactedlikewise.withitsactionantagonizablebyspantide,aSPreceptorblocker.TofurthercharacterizethenatureofcellsresponsivetothechallengeofSP,immunocytochemicalstainingagainstS-100proteinandsomeadenohypophysealhormoneswasperformedaloneorplusautoradiography.TheresultsshowedthatthepercentageofS-100proteinimmunorectivecellswasapparentlyelevatedbytheaddtionofNle^11-Spfor48h,whichindicatesapreferentialproliferationoffolliculo-stellatecellsundertheregime.ThiswasconfirmedbyincreasesinimmunocytochemicalorautoradiographicallabellingindicesofanteriorpituitarySubstancePandanteriorpituitarycellproliferation.Cellstreatedsimilarly.Takentogether,TheseresultsrevealthatthetrophicactionofSPobservedpreviouslyinothertissuesisalsopresentatleastinculturedratanteriorpituitarycells.withrespondingcellsbeingpredominantlyfolliculo-stellatecellsastypifiedbyS-100proteinimmunoreactivity.Therefore,anintra-pituitarytrophicactionofSPinvivocouldbeanticipated.
简介:cAMPmediatedsignalingmayplayasuppressiveroleinimmuneresponse.WepreviouslyfoundthatthecAMP-elevators(CTxand8-Br-cAMP)inhibitedIL-12,IL-la,IL-6geneexpression,butincreasedthetranscriptionallevelsofIL-10andIL-1RainLPS-treatedmurineperitonealmacrophages.ThepresentstudyexaminedapossiblemolecularmechanisminvolvedincAMPelevators-inducedinhibitionofIL-12p40expressioninresponsetoLPS.OurdatademonstratedthatcAMPelevatorsdownregulatedIL-12p40mRNAexpressionandIL-12pT0productioninmurineperitonealmacrophages.SubsequentstudiesrevealedthatcAMP-elevatorsblockedphosphorylationofp38MAPK,butdidnotaffecttheactivityofNF-κBbindingtoIL-12promoter(-136/-112).ThisisthefirstreportthatcAMPelevatorsinhibitLPS-inducedIL-12productionbyamechanismthatisassociated,atleastinpart,withp38-dependentinhibitionbycAMPsignalingpathways.
简介:TherearetwopossibleoutcomeswhenDNAdamageoccursinnormalmammaliancells:eitherinductionofcell-cyclecheckpointwhichinhibitstheprogressofthecellcyclesaswellasactivatesDNArepairpathways,oractivationofapoptosistoeliminatedamagedcells.Thep53tumour-suppressorgeneplaysakeyroleinselectingthesepathways.Inourpresentworks,thehumangastriccancercelllineAGSwastreatedwithtripchlorolide,apotentantitumorcompoundpurifiedfromaChineseherbTripterygiumWilfordiiHook.Singlecellgelelectrophoresis(Cometassay)showedthatthetreatmentoftripchlorolideresultedinDNAdamageinAGScells.ThedamagedAGScellswentthroughapoptosis,whichwastime-anddose-dependent.
简介:从胖身体和棉花一种蛾的幼虫的中间的勇气的细胞色素P450(Cyt-P450)蛋白质,Helicoverpa有佩带徽章权利的人一,被一套纯化过程部分分别地包括微分离心分离,小伙子的增溶,由木钉降水的蛋白质降水和DE-32柱层析净化。Cyt-P450被COdifference光谱和SDS页的方法检测。从胖身体ofH的净化的solubilized微体的部分。有佩带徽章权利的人一多于17褶层被净化。三个蛋白质乐队被SDS页与分子的群众检测70600,63300和571200Da。有63300和571200Da的分子的质量的蛋白质是Cyt-P450的isozymes,是可能的。
简介:Recentstudiesindicatethatcell-cyclecheckpointsaretightlycorrelatedwiththeregulationofapoptosis,inwhichp53playsanimportantrole.OurpresentworksshowthattheexpressionofE6/E7oncogenesofhumanpapillomavirusinHeLacellsisinhibitedinthepresenceofanti-tumorreagenttripchlorolide(TC),whichresultsintheup-regulationofp53inHeLacells.Interestingly,underthesameTC-treatment,thecellsattheearlyS-phasearemoresusceptibletoapoptosisthanthoseatthemiddleS-phasealthoughp53proteinisstabilizedtothesamelevelinbothsituations.Significantdifferenceisexhibitedbetweenthetwospecifiedexpressionprofiles.Furtheranalysisdemonstratesthatanti-apoptoticgenesurvivinisup-regulatedbyp53intheTC-treatedmiddle-Scells,whereasitisdown-regulatedbyp53intheTC-treatedearly-Scells.Takentogether,thepresentstudyindicatesthatthedifferentialp53-regulatedexpressionofsurvivinatdifferentstagesofthecellcycleresultsindifferentcellularoutputsunderthesameapoptosis-inducer.
简介:CREB有约束力的蛋白质(CBP)和它的相当或相同事物p300是涉及细胞的活动的一个宽数组的各种各样的顺序特定的抄写因素的transcriptionalco使活跃之物,例如脱氧核糖核酸修理,房间生长,区别和apoptosis。Severalstudies建议了CBP和p300可能被看作瘤压制ors,与他们是的突出的角色响应各种各样的刺激的不同基因表示模式的跨coupling。他们主要经由乙酰化施加行动嘘和另外的规章的蛋白质(例如p53)。在CBP/p300功能的主要悖论是他们似乎能够贡献各种各样的反对细胞的进程。呼吸上皮tumorigenesis代表一个范围的多步累积的一个复杂过程基因并且epigenetic错误。通过激活和压抑的建筑群的交替的形成的Transcriptionmodulation是这些精神错乱的最终的收敛点,并且CBP/p300在这相互影响代表关键参加者。因此,他们的分子的行动和相互作用的照明能在呼吸上皮carcinogenesis为药理学干预揭示新潜在的目标。
简介:Amajorproblemwhichispoorlyunderstoodinthemanagementofbladdercancerislowsensitivitytochemotherapyandhighrecurrenceaftertransurethralresection.Insulin-likegrowthfactor1receptor(IGF-1R)signalingplaysaveryimportantroleinprogression,invasionandmetastasisofbladdercancercells.Inthisstudy,weinvestigatedwhetherIGF-1Rwasinvolvedinthegrowthstimulatingactivityanddrugresistanceofbladdercancercells.Theresultsshowed:ThemRNAsofIGF-1,IGF-2andIGF-1Rwerestronglyexpressedinserum-freeculturedT24cellline,whereasnormalurothelialcellsdidnotexpressthesefactors/receptorsoronlyintracelevels;T24cellrespondedfarbettertogrowthstimulationbyIGF-1thandidnormalurothelialcells;blockageofIGF1Rbyantisenseoligodeoxynucleotide(ODN)significantlyinhibitedthegrowthofT24cellandenhancedsensitivityandapoptosisofT24cellstomitomycin(MMC).TheseresultssuggestedthatblockageofIGF-IRsignalingmightpotentiallycontributetothetreatmentofbladdercancercellswhichareinsensitivetochemotherapy.
简介:目的:通过检测藏獒黑素皮质激素受体1(MClR)基因的单链构象多态性(SSCP)在不同毛色群体中的分布,探讨MClR基因多态性与毛色表型的相关性。方法:采用DNA测序技术,选择不同毛色藏獒的DNA为样本,根据GenBank发布的荷斯坦牛MClR基因序列设计一对引物,采用PCR-SSCP技术分析MClR基因在藏獒中的SSCP。结果:MClR基因在藏獒中具有PCR-SSCP多态性,分别检测到3种基因型(AA、AB和BB);对MClR基因多态性片段DNA克隆测序后发现,MClR基因在编码区第313位存在单碱基突变(G-,A),该突变导致第105位氨基酸发生由丙氨酸向苏氨酸的改变(T105A)。结论:肘CJR基因的多态性与毛色性状不存在显著的相关性。
简介:Changesinthedistributionof1P1-antigeninthedevelopingchickretinahavebeenexaminedbyindriectimmunofluorescencestainingtechniqueusingthenovelmonoclonalantibody(MAb)1P1.Expressionofthe1P1antigenwasfoundtoberegulatedinradialaswellasintangentialdimensionoftheretina,beingpreferentiallyorexclusivelylocatedintheinnerandouterplexiformlayersoftheneuralretinadependingonthestagesofdevelopment,Withtheonsetoftheformationoftheinnerplexiformlayer1P1antigenbecomesexpressedintheretina.Withprogressingdifferentiationoftheinnerplexiformlayer1P1immunofluorescencerevealed2subbandsatE9and6subandsatE18,Atpostnatalstages(afterP3)immunoreactivitywasreducedinaninside-outsidesequenceleadingtothecompleteabsenceofthe1P1antigeninadulthood.1P1antigenexpressionintheouterplexiformlayerwasalsosubjecttodevelopmentalregulation.Thespation-temporalpatternof1P1antigenexpressionwascorrelatedwiththetimecourseofhistologicaldifferentationofchickretina,namelythesynapserichplexiformlayers.Whetherthe1P1antigenwasfunctionallyinvolvedindendriteextensionandsynapseformationwasdiscussed.