摘要
应用RT PCR技术 ,从分泌抗隐孢子虫表膜单克隆抗体 (McAb)的杂交瘤细胞 4G4中 ,扩增出抗体VH 和VL 基因 ,用Linker (Gly4 Ser) 3基因 ,将VH 和VL 基因连接成ScFv基因 ,并将其克隆至pMD 18T载体中。经核甘酸序列分析证实 ,VH 和VL 基因及Linker基因拼接正确 ,基因全长 717bp ,经计算机分析 ,VH和VL 基因均为新发现的基因序列 。
The V H and V L genes were amplified from a hybirdoma cell line 4G4 producing mouse McAb against Cryptosporidium parvum sporozoite membrane by RT-PCR.The V H and V L genes were connected through a flexible linker(Gly 4Ser) 3 and the V H-linker-V L(ScFv)fusion gene was cloned into c clone vector pMD18-T.The ScFv gene was sequenced,analyzed by computer.The ScFv gene consists of 717bp encoding 239 aminoacid residues.Both V H and V L genes were reconfirmed as functionally rearranged mouse immunoglobulin variable region genes and appeared to be new genes.
出处
《寄生虫与医学昆虫学报》
CAS
2004年第4期198-202,共5页
Acta Parasitologica et Medica Entomologica Sinica
基金
国家自然科学基金资助 (编号 3 9970 5 63 )
