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OALib Journal期刊
ISSN: 2333-9721
费用：99美元
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人gfp-awp1融合基因载体的构建及其在293细胞中的表达
Keywords: awp1,绿色荧光蛋白,293细胞
Abstract:
目的克隆、构建绿色荧光蛋白(gfp)-awp1(associatedwithproteinkinasecrelatedkinase1,awp1)表达载体,观察awp1在293细胞中表达和定位。方法采用逆转录pcr(rt-pcr)法从人ecv304内皮细胞中扩增awp1cdna编码区,并将其重组于gfp表达载体pegfp-c2中。经酶切、序列鉴定分析后,将该重组质粒通过dotap脂质体介导,转染293细胞。荧光显微镜观察awp1在细胞内的表达和分布。结果gfp-awp1融合基因表达载体经酶切鉴定和测序分析确认构建成功,并在293细胞中获得了高效表达。荧光显微镜下,在不携带外源基因的空载体pegfp-c2转染的对照组293细胞,绿色荧光均匀分布于整个细胞中;在重组质粒pegfp-c2/awp1转染的293细胞,绿色荧光弥散分布于细胞质内。结论成功构建gfp-awp1融合基因表达载体并表达于293细胞胞质中。
| [1] | tsienry.thegreenfluorescentprotein[j].annurevbiochem,1998,67,509-44.
|
| [2] | duanw,sunb,litw,etal.cloningandcharacterizationofawp1,anovelproteinthatassociateswithserine/threoninekinaseprk1invivo[j].gene,2000,256(1-2):113-21.
|
| [3] | 莫永炎,曹永宽,刘亚伟,等.人his-awp1融合蛋白表达载体的构建及其在原核生物的表达[j].中华神经医学杂志,2003,2(2):123-5.moyy,caoyk,liuyw,etal.constructionofhis-taghumanawp1fusionproteinexpressionvectoranditsexpressioninprokaryoticcells[j].chinjneuromed,2003,2(2):123-5.
|
| [4] | jiangy,liuah,huangqb,etal.p38mapksignalisnecessaryfortnf-αgeneexpressioninrawcells[j].actabiochimbiophyssin,1999,31(1):9-15.
|
| [5] | amanom,mukaih,onoy,etal.identificationofaputativetargetforrhoastheserine-threoninekinaseproteinkinasen[j].science,1996,271(5249):648-50.
|
| [6] | watanabeg,saitoy,madaulep,etal.proteinkinasen(pkn)andpkn-relatedproteinrhophilinastargetsofsmallgtpaserho[j].science,1996,271(5249):645-48.
|
| [7] | kluga,schwabejw.proteinmotifs5.zincfingers[j].fasebj,1995,9(8):597-604.
|
| [8] | kurodas,tokunagac,kiyoharay,etal.protein-proteininteractionofzincfingerlimdomainswithproteinkinasec[j].jbiolchem,1996,271(49):31029-32.
|
| [9] | zhangl,jiangy.theintracellularlocalizationofp38mapkinaseindifferentprimaryculturedcells[j].actabiophyssin,2000,16(3):481-8.
|
| [10] | jiangy,gongxw.regulationofinflammatoryresponsesbymapksignaltransductionpathways[j].actaphysiolsin,2000,52(4):267-71.
|
| [11] | hoodjk,silverpa.inorout?regulatingnucleartransport[j].curropincellbiol,1999,11(2):241-7.
|
| [12] | hoodjk,silverpa.diversenucleartransportpathwaysregulatecellproliferationandoncogenesis[j].biochimbiophysacta,2000,1471(1):m31-41.
|
| [13] | jansda,xiaocy,lammh.nucleartargetingsignalrecognition:akeycontrolpointinnucleartransport[j]?bioessays,2000,22(6):53244.
|
| [14] | dingwallc,laskeyra.nucleartargetingsequences-aconsensus[j]?trendsbiochemsci,1991,16(12):478-81.
|
| [15] | kalderond,robertsbl,richardsonwd,etal.ashortaminoacidsequenceabletospecifynuclearlocation[j].cell,1984,39(3pt2):499-509.
|
| [16] | cingolanig,petosac,weisk,etal.structureofimportin-betaboundtotheibbdomainofimportin-alpha[j].nature,1999,399(6733):221-9.
|
| [17] | izaurraldee,kutayu,vonkobbec,etal.theasymmetricdistributionoftheconstituentsoftheransystemisessentialfortransportintoandoutofthenucleus[j].emboj,1997,16(21):6535-47.
|
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