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OALib Journal期刊
ISSN: 2333-9721
费用：99美元
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cnksr2基因干扰腺病毒的构建及鉴定
, PP. 103-107
Keywords: cnksr2基因,干扰,腺病毒
Abstract:
构建小鼠cnksr2基因干扰腺病毒质粒载体并加以鉴定.根据小鼠cnksr2基因序列设计cnksr2干扰序列引物,定向克隆至穿梭载体pshuttle-h1的bglⅱ和hindⅲ位点,pmeⅰ酶切线性化的pshuttle-h1-sicnksr2干扰质粒,并与腺病毒载体(padeasy-1质粒)共同转化e.colibj5183感受态细菌,产生重组腺病毒载体.用pacⅰ酶切线性化的回收质粒,转染293a细胞,包装腺病毒颗粒,在倒置显微镜下观察细胞病理性变化(cellpathologicaleffect,cpe),用tcid50法测定病毒颗粒的浓度,并初步观察病毒感染pc12细胞对目的基因的干扰效率.经酶切鉴定、测序证实成功构建小鼠cnksr2基因干扰腺病毒载体.包装的腺病毒浓缩悬液滴度为3.98×107pfu/ml.cnksr2在大鼠及小鼠中具有保守性,该病毒能在大鼠来源的pc12细胞中成功表达,并且对cnksr2基因干扰效率达50%以上.结论:成功构建了小鼠cnksr2基因的干扰腺病毒载体.
| [1] | yaoi,hatay,iden,etal.maguin,anovelneuronalmembrane-associatedguanylatekinase-interactingprotein[j].jbiolchem,1999,274(17):11889-11896.
|
| [2] | royf,iabergeg,douziechm,etal.ksrisascaffoldrequiredforactivationoftheerk/mapkmodule[j].genesdev,2002,16(4):427-438.
|
| [3] | bumeisterr,rossec,anselmoa,etal.cnk2couplesngfsignalpropagationtomultipleregulatorycascadesdrivingcelldifferentiation[j].currbiol,2004,14(5):439-445.
|
| [4] | santossd,verveerpj,bastiaenspi.growthfactor-inducedmapknetworktopologyshapeserkresponsedeterminingpc-12cellfate[j].natcellbiol,2007,9(3):324-330.
|
| [5] | vaudryd,storkpj,lazarovicip,etal.signalingpathwaysforpc12celldifferentiation:makingtherightconnections[j].science,2002,296(5573):1648-1649.
|
| [6] | pearsong,robinsonf,beersgibsont,etal.mitogen-activatedprotein(map)kinasepathways:regulationandphysiologicalfunctions[j].endocrrev,2001,22(2):153-183.
|
| [7] | therrienm,wongam,kwane,etal.functionalanalysisofcnkinrassignaling[j].procnatlacadsciusa,1999,96(23):13259-13263.
|
| [8] | douziechm,sahmim,labergeg,etal.aksr/cnkcomplexmediatedbyhyp,anovelsamdomain-containingprotein,regulatesras-dependentrafactivationindrosophila[j].genesdev,2006,20(7):807-819.
|
| [9] | roignantjy,hamels,janodyf,etal.thenovelsamdomainproteinaveugleisrequiredforrafactivationinthedrosophilaegfreceptorsignalingpathway[j].genesdev,2006,20(7):795-806.
|
| [10] | 卢大儒,邱信芳.腺病毒载体、腺病毒相关病毒载体与基因治疗[j].国外医学遗传学分册,1995,18:61-64.
|
| [11] | 何金生,王健伟,涛洪.腺病毒载体构建原理与方法的研究进展[j].中华实验和临床病毒学杂志,2001,15:399-400.
|
| [12] | therrienm,wongam,rubingm.cnk,araf-bindingmultidomainproteinrequiredforrassignaling[j].cell,1998,95(3):343-353.
|
| [13] | nagaset,ischikawak,suyamam,etal.predictionofthecodingsequencesofunidentifiedhumangenes.？.thecompletesequencesof100newcdnaclonesfrombrainwhichcodeforlargeproteinsinvitro[j].dnares,1998,5(6):355-364.
|
| [14] | ohtakarak,nishizawam,izawai,etal.densin-180,asynapticprotein,linkstopsd-95throughitsdirectinteractionwithmaguin-1[j].genescells,2002,7(11):1149-1160.
|
| [15] | eastwoodsl,burnetpw,harrisonpj.alteredsynaptophysinexpressionasamarkerofsynapticpathologyinschizophrenia[j].neuroscience,1995,66(2):309-319.
|
| [16] | deakinjf,simpsonmd.atwo-processtheoryofschizophrenia:evidencefromstudiesinpost-mortembrain[j].jpsychiatrres,1997,31(2):277-295.
|
| [17] | hougeg,rasmussenih,hovlandr.loss-of-functioncnksr2mutationisalikelycauseofnon-syndromicx-linkedintellectualdisability[j].molsyndromol,2012,2(2):60-63.
|
| [18] | friedmana,perrimonn.afunctionalrnaiscreenforregulatorsofreceptortyrosinekinaseanderksignalling[j].nature,2006,444(7116):230-234.
|
| [19] | anselmoan,burneisterr,thomasjm,etal.criticalcontributionoflinkerproteinstorafkinaseactivation[j].jbiolchem,2002,277(8):5940-5943.
|
| [20] | yaoi,ohtsuket,kawabeh,etal.associationofmembrane-associatedguanylatekinase-interactingprotein-1withraf-1[j].biochembiophysrescommun,2000,270(2):538-42.
|
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