胃癌相关基因GCRG213在真核细胞的定位研究  

Location of gastric cancer related gene GCRG213 in eukaryotic cells

作　　者：高利利[1] 王孟薇[1] 吴本俨[1] 周涛[2] 黄海力[1] 尤纬缔[1] 王卫华[1] 

机构地区：[1]解放军总医院南楼消化科,北京100853 [2]军事医学科学院仪器中心,北京100853

出　　处：《中华肿瘤防治杂志》2007年第2期90-92,103,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(20370635)

摘　　要：目的研究胃癌相关基因GCRG213在真核细胞的定位。方法采用聚合酶链反应(PCR)技术,从pGEM-T质粒上选择性扩增出包含完整ORF在内的人胃癌相关基因GCRG213的DNA片段,将目的片段两端分别引入限制性内切酶PstI和BamHI识别位点,克隆至真核表达载体pEGFP-Nl。将测序正确的阳性重组子pEGFP-Nl-GCRG213采用脂质体瞬时转染COS-7细胞,激光共聚焦技术检测GCRG213基因在真核细胞内的蛋白定位。结果经测序证实,GCRG213正确克隆入真核表达载体pEGFP-Nl,组成阳性重组子pEGFP-Nl-GCRG213。测序正确的pEGFP-N1-GCRG213重组子经脂质体转染COS-7细胞,激光共聚焦显微镜观察GCRG213蛋白在胞核和胞质中均有表达。结论GCRG213蛋白在胞核和胞质中均有表达。OBJECTIVE： To study the location of gastric cancer related gene GCRG213 in eukaryotic cells. METHODS： GCRG213＇s DNA fragments with complete open reading frames were amplified by PCR from plasmid pGEM-T, and then were cloned into eukaryotic expression vector pEGFP-N1 after introducing the sites of restrictive endonuclease enzyme Pst I and BamH I, transiently transfecting the eukaryotic cell COS-7 with the recombinant plasmid pEGFP-N1-GCRG213, and applying eonfocal microscopy to observe the location of transfected GCRG213 in COS-7 cell. RESULTS： DNA sequence analysis showed that the GCRG213 sequence was right in the recombinant plasmid pEG- FP-N1-GCRG213, transiently transfeeting the eukaryotic cell COS-7 with pEGFP-N1-GCRG213. By using the confocal technique, GCRG213 was identified to be expressed in the cytoplasm and nuclei of COS-7 cells. CONCLUSION： Transient transfection shows that GCRG213 is located in cytoplasm and nuclei of COS-7 cells.

关 键 词：胃肿瘤/遗传学 多态性 限制性片段长度 基因表达 转染 聚合酶链反应 

分 类 号：R735.2[医药卫生—肿瘤]