摘要
目的 构建含人 MUC- 1全长 c DNA序列的真核表达载体 ,并观察其在 COS- 7细胞中的表达 .方法 将目的基因 MUC- 1克隆入 p GEM- 3zf(- )载体中并进行酶切鉴定和DNA序列测定 .亚克隆入真核质粒 pc DNA3.1(+) ,构建真核表达载体 pc DNA3.1(+) - MU C- 1.用电穿孔法将重组质粒转入 COS- 7细胞 ,以免疫荧光和流式细胞仪检测 MUC- 1的表达 .结果 酶切鉴定和序列分析证实 ,重组质粒含有人MU C- 1全长 c DNA编码序列 ,转染实验表明 MU C- 1基因能在 COS- 7细胞中表达 .结论 人 MU C- 1全长 c DNA基因真核表达载体构建及其在 COS- 7中的表达均获成功 。
AIM To construct an eukaryotic expression vector containing the coding region of human full length MUC1 gene and to detect its expression in COS 7 cells.METHODSThe MUC1 gene was ligated into the sites of HindIII and Sal I of cloning vector pGEM 3zf. After identification, the gene was subcloned into eukaryotic expression vector pcDNA3.1(+). The recombinant pcDNA3.1(+) MUC 1 was transfected into COS 7 cells by electroporation. Immunoflourescence and FCM were used to detect the MUC1 gene expression in COS 7 cells. RESULTS Restriction analysis and DNA sequencing showed that the recombinant plasmid contained the coding region of human full length MUC1 gene. Transfection experiment verified that MUC1 gene could be expressed in COS 7 cells. CONCLUSION The eukaryotic expression vector containing the MUC1 gene was succesfully constructed and expressed, which is a basis for the study for DNA vaccine.
出处
《第四军医大学学报》
北大核心
2002年第13期1231-1234,共4页
Journal of the Fourth Military Medical University
