【摘要】 采用巢式PCR(nested PCR)的方法从Raji细胞中扩增出人CD80 cDNA,并将这一cDNA克隆入载体pUC19中。经PCR扩增和限制性酶切分析,进行初步鉴定后,再将人CD80 cDNA克隆到pcD-NA表达载体上,构建成为pCD-CD80重组质粒。应用脂质体介导的基因转移技术将这一表达载体导入小鼠黑色素瘤B16细胞后,用SABC法进行瞬时表达的检测。结果表明,转染后48h就具有明显人CD80基因的表达产物。更多还原

【Abstract】 Human CD80 cDNA was amplified by nested PCR method from the CD80+ Raji cells and cloned to vector pUC19. The cDNA fragment was identified by enzyme digestion and PCR detection. Then a pCD-CD80 recombinant plasmid was constructed by cloning human CD80 cDNA into expression vector pcD-NA. Melanoma B16 cells of mice were transfected with pCD-CD80 plasmid by lipofectin mediated gene transfer, the transient expressive results were detected by SABC method. It was observed that the expression of human CD80 gene was detected 48 h after transfecting distinctly.更多还原