简介:Changesinthedistributionof1P1-antigeninthedevelopingchickretinahavebeenexaminedbyindriectimmunofluorescencestainingtechniqueusingthenovelmonoclonalantibody(MAb)1P1.Expressionofthe1P1antigenwasfoundtoberegulatedinradialaswellasintangentialdimensionoftheretina,beingpreferentiallyorexclusivelylocatedintheinnerandouterplexiformlayersoftheneuralretinadependingonthestagesofdevelopment,Withtheonsetoftheformationoftheinnerplexiformlayer1P1antigenbecomesexpressedintheretina.Withprogressingdifferentiationoftheinnerplexiformlayer1P1immunofluorescencerevealed2subbandsatE9and6subandsatE18,Atpostnatalstages(afterP3)immunoreactivitywasreducedinaninside-outsidesequenceleadingtothecompleteabsenceofthe1P1antigeninadulthood.1P1antigenexpressionintheouterplexiformlayerwasalsosubjecttodevelopmentalregulation.Thespation-temporalpatternof1P1antigenexpressionwascorrelatedwiththetimecourseofhistologicaldifferentationofchickretina,namelythesynapserichplexiformlayers.Whetherthe1P1antigenwasfunctionallyinvolvedindendriteextensionandsynapseformationwasdiscussed.
简介:Arabidopsisthaliana嘘一deacetylase1(AtHD1或AtHDA19),酵母RPD3的一个相当或相同的事物,是在植物的许多生理、发展的过程的一个全球管理者。尽管有为在植物基因规定和开发的AtHD1的一个角色的基因证据,AtHD1的生物化学、细胞的性质糟糕被理解。这里,我们在vivo报导AtHD1的细胞的本地化模式并且嘘在vitro的一项deacetylase活动。绿荧光灯的蛋白质(GFP)的短暂、稳定的表示在洋葱房间标注了AtHD1并且分别地,在转基因的Arabidopsis的根,种子和叶子表明AtHD1在euchromatic区域大概在原子核是局部性的并且从核排除。AtHD1的本地化模式与涉及核形成和transgenes的silencing并且分别地重复了DNA元素的AtHD2和AtHDA6的那些不同。另外,一hist一deacetylase活动试金证明在细菌生产的recombinantAtHD1示威了一特定嘘在vitro的一项deacetylase活动。数据建议AtHD1是原子蛋白质并且拥有嘘为对植物生长和开发重要的全球transcriptional规定负责的一项deacetylase活动。
简介:用原子力显微镜(AFM)观察线性DNA并探讨其成像条件。用RT-PCR技术扩增柯萨奇B1病毒VP1基因DNA片段,纯化回收后配制成含和不含1mmol/LMgCl2的水溶液,DNA终浓度为100μg/ml。分别取20μl滴加在新鲜解理的云母片上,吸收1min,用滤纸吸去残液,氮气吹干,在室温下采用MultiModeAFMNanoscopeⅢa的敲击模式成像。同时比较新制备和多次使用过的探析所获得图像的质量,对两种探针进行扫描电镜观察。电泳证明获得了0.83kb的VP1基因DNA线性片段,Mg^2+存在时,DNA在云母片上吸附延展较好,所获图像质量优于无Mg^2+时。新制备的探针较多次使用过的探针所获图像分辨率更高。DNA分子的表现宽度为18±2.9nm,高度为0.8±0.2nm。说明AFM能以高分辨率直接观察DNA分子,Mg^2+的存在和高质量的探针有助于获得理想的图象。
简介:TheyeastHAL1genewasintroducedintoArabidopsisthalianabyAgrobacteriumtumefaciens-mediatedtransformationwithvacuuminfiltrationunderthecontrolofCaMV35Spromoter.Thirty-threeindividualkanamycinresistantplantswereobtainedfrom75,000seeds.SouthernblottinganalysisindicatedthatHAL1genehadbeenintegratedintoallofthetransgenicplants'genomes.ThecopynumberofHAL1geneintransgenicplantswasmostly1to3bySouthernanalysis.Phenotypesoftransgenicplantshavenodifferenceswithwildtypeplants.Severalsamplesoftransformantswereself-pollinated,andprogeniesfromtransformedandnon-transformedplants(controls)wereevaluatedforsalttoleranceandgeneexpression.MeasurementofconcentrationsofintracellularK+andNa+showedthattransgeniclineswereabletoretainlessNa+thanthatofthecontrolundersaltstress.ResultsfromdifferenttestsindicatedtheexpressionofHAL1genepromotesahigherlevelofsalttoleranceinvivointhetransgenicArabidopsisplants.
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.
简介:SomerecentstudiesindicatedthatGABAergicsystemisinvolvedinmammalianspermacrosomereaction(AR),butdirestevidencepertainingtotheexpressionofgatlinmammalianspermisnotyetdemonstrated.Inthisstudy,weevaluatedthepresenceof67kDaGAT1proteinandmRNAinrattestisbyWesternblottingandreversetranscription-polymerasechainreaction.Meanwhile,immunohistochemicalandimmunofluorescentanalysesalsoidentifiedGAT1proteinontheelongatedspermatidandsperm.Theseresultsindicatedthatrattestisisanovelsiteofgatlexpression.FurtherstudiesshouldbetakentoexploretheroleofGAT1proteinonspermacrosomereaction.
简介:以便学习TGF-beta1的结构功能细节,重组体老鼠TGF-beta1的成熟形式在细菌被表示。TGF-beta1(氨基酸279-390)的112氨基酸的carboxyl终端部分的合成被一个可诱导的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的活跃、选择的合成。在减少条件下面在SDS-polyacrylamide胶化上决定的表示TGF-alpha1单体的分子量是大约13kD。连续净化洗与纯化步是足够的净化表示产品到同质的单个胶化过滤结合了。氨基终端的定序表明重组体蛋白质的N终端与出版数据相同。在西方的污点分析,重组体多肽对polyclonalTGF-beta1抗体显示出优秀antigenicity。在这研究表示的成熟重组体老鼠TGF-beta1提供一个有用工具因为未来详细说明了结构、功能的研究。
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.
简介:尽管许多进步在理解花的机关身份怎么在花的发展期间是坚定的被取得了,更少花的机关怎么在晚花的发展阶段被详细描述被知道。这里,我们描述新奇花的异种,起皱的花瓣和stamens1(wps1),它在花瓣和雄蕊的发展显示出缺点。基因分析显示那wps1异种相应于在染色体3的长手臂的一个单个后退的地点。在wps1异种的花的机关的早发展在野类型类似于那,并且异种的malfunction在晚发展的阶段开始,显示花瓣和雄蕊的外观上的一个缺点。在成熟的花,在异种的花瓣和雄蕊细丝被起皱或合拢,并且在花瓣和雄蕊细丝的L1层下面的细胞的形态学是反常的。花的机关身份基因的表达式模式没与野类型的相比在wps1异种被影响,这被发现,与所有花的机关的未改变的开发一致。而且,在花瓣的不同类型的表皮的房间的身份被维持。组织学的分析证明在wps1花,所有花瓣不规则地被合拢,并且在花瓣有系的结构,当内部房间的形状和安排畸形、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花的机关身份基因,并且工作在迟了的花期间调制细胞的区别发展阶段。
简介:GABAtransporter1(GAT1)takesimportantrolesinmultiplephysiologicalprocessesthroughtheuptakeandreleaseofGABA,buttheregulationofGAT1geneexpressionindifferenttissuesisrarelyknown.Toaddressthequestion,first,5'RapidamplificationofcDNAend(RACE)wasusedtodetermineGAT1transcriptionalstartingsitesinneonatalmousecerebralcortexandintestine,adultmousebrainandadultrattestis.Theproductsof5'RACEwereconfirmedbyDNAsequencing.WefoundthatthetranscriptofGAT1inneonatalmousecerebralcortexandadultmousebrainstartsatthesamesite(insideofexon1),whileinmouseintestine,GAT1startstranscriptioninintron1,andinrattestis,thetranscriptofGAT1hasanadditionaluntranslationexontothe5'direction.