抗CD_3单链抗体基因克隆与表达

Clone and Expression of Anti-CD_3 Single Chain Antibody Gene

采用重叠延伸拼接法 (SOE)连接抗CD3 抗体的重链可变区 (VH)和轻链可变区 (VK)基因 ,构建单链抗体基因 (ScFv)并将其克隆于含Tac启动子质粒中 ,用斑点杂交法筛选出重组子 ,转化大肠杆菌BL2 1(DE3 )

The genes at the variable regions of heavy (V H) and light (V K) chains of anti CD 3 antibody were linked by splicing, overlapping and extension(SOE). The single chain antibody (ScFv) gene constructed in this way was cloned into the plasmid containing Tac promotor, then recon was selected out by dot blot hybridization and transformed to E. coli BL 21 (DE3). The result showed that anti CD 3 single chain antibody protein was successfully expressed.