摘要
目的:研究gp130胞内区的假定核定位序列(nuclearlocalizationsequence,NLS)是否具有核定位功能。方法:首先将编码SV40大T抗原NLS(阳性对照)和gp130胞内区假定NLS的cDNA分别插入真核表达载体pEGF-PN1,构建成表达质粒pSVNLSGFP和pGPNLSGFP。然后将这两种表达质粒分别转染HeLa细胞并采用genecitin筛选稳定表达细胞株。以加强型绿色荧光蛋白(enhancedgreenfluorescenceprotein,EGFP)在细胞内的分布情况作为观察指标确定融合蛋白表达产物在细胞内的定位。结果:稳定转染表达质粒pSVNLSGFP的HeLa细胞中绿色荧光主要集中分布在细胞核内;而转染了pGPNLSGFP的HeLa细胞中绿色荧光在细胞浆和细胞核内呈现出均匀分布状态。结论:gp130胞内区的假定NLS序列不具有核定位功能。
Objective：This stud y was designed to investigate whether the putative nuclear localization sequence （NLS）located in the intracellular domain of gp１３０possesses the function of nuclear import．Methods ：The expression plasmids pSV－NLS－GFP and pGP－NLS－GF P，constructed by inserting the cDNA coding S V４０large T－antigen NLS（as posi tive control）and the putative gp１３０NLS into the ve ctor pEGFPN１，were intro duced into HeLa cells and the stable－transfecta nts were selected in the presen ce of genecitin．Subcellular localization of the f usion protein was identified by enhanced green fluorescence protein（EGFP）．Results：Green fluorescence accu mulated in t he nucleus of HeLa cells transfected with pSV－NLS－GFP，but shared equally in t he cytoplasm and nucleus in pGP－NLS－GFP transfected HeLa cells． Conclusion：The target NLS in gp１３０doesn't posse ss the function of nuclear i mport．
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2001年第10期1029-1032,共4页
Chinese Journal of Cancer
