肿瘤相关抗原HCA520真核表达载体的构建及细胞内表达的定位分析  

Construction of HCA520 and GFP Fusion Gene Eukaryotic Expression Recombinant and Analysis Its Cell Location

作　　者：杨美香[1,2] 曲迅[1] 韩克军[2] 马道新[1] 陈慰峰[2] 

机构地区：[1]山东大学齐鲁医院临床基础研究所,济南250012 [2]北京大学医学部免疫学系,北京100083

出　　处：《细胞生物学杂志》2004年第2期193-196,共4页Chinese Journal of Cell Biology

基　　金：国家"973"基金资助项目(No.G1999053904);北京市自然科学基金资助项目(No.7001002)~~

摘　　要：应用PCR技术获得肿瘤相关抗原HCA520编码基因,构建至pGEM-T Easy载体,测序正确后,亚克隆至pEGFP-N1载体,1％的琼脂糖电泳得到两条带,与预期结果相同;鉴定正确的基因一过性转染至CHO细胞,共聚焦显微镜观察HCA520基因编码蛋白主要位于胞浆靠近胞膜处,为HCA520功能的进一步研究打下了基础。Using PCR technique to obtain HCA520 encoding gene and cloned into the pGEM-T Easy vector. The fusion gene was identified by DNA sequencing. The identified gene was subcloned into the pEGFP-N1 vector and further identified by Sall and XhoI restriction enzymes. The confirmed recombination was transfected into the CHO cells, analysis the cell location using laser scanning confocal microscope. Most of the HCA520 protein was located in the cytoplasm. The eukaryotic expression recombination of HCA520 and its cell location analysis were valuable for further study of HCA520.

关 键 词：肿瘤 抗原 HCA520 真核表达载体 细胞定位 

分 类 号：R392[医药卫生—免疫学]