摘要
应用 RT-PCR技术 ,从两株分泌具有中和活性的抗 A型产气荚膜梭菌 α毒素单克隆抗体 ( Mc Ab)的杂交瘤细胞株 2 E3和 1 A8中 ,分别扩增出抗体 VH 和 VL 基因 ,用 Linker( Gly4Ser) 3基因 ,将 VH 和 VL 基因连接成Sc Fv基因 2 E3-Sc Fv和 1 A8-Sc Fv,并将其克隆至 p GEM-T载体中 .经核苷酸序列分析证实 ,VH 和 VL 基因以及Linker基因拼接正确 ,2 E3-Sc Fv基因全长为 72 9bp,经计算机分析 ,VH 和 VL 基因均为新发现的基因序列 ,符合功能性重排的鼠抗体可变区基因特征 .2 E3-Sc Fv的 VH 和 VL 基因分别属于鼠免疫球蛋白重链 ( B)和轻链κ 家簇 ;而 1 A8-Sc Fv的 VH 和 VL 基因分别属于鼠免疫球蛋白重链 ( A)和轻链 κ
The V H and V L genes were amplified from two hybridoma cell lines producing mouse McAb against alpha txoin of Clostridium perfringens type A 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 a clone vector pGEM T.The 2E3 ScFv and 1A8 ScFv gene were sequenced and analyzed by computer.The ScFv genes consist of 726bp and 729bp respectively.Both V H and V L genes were functionally rearranged mouse immunoglobulin variable region genes and appeared to be new genes.According to Kabat classed method,the V H gene segment and V L gene segment of 2E3 ScFv belong to the mouse Ig heavy chain subgroupⅡ(B) and κ chain subgroup Ⅲ respectively,but the V H gene segment and V L gene segment of 1A8 ScFv belong to the mouse Ig heavy chain subgroup Ⅱ(A) and κ chain subgroup Ⅵ respectively.
出处
《河北师范大学学报（自然科学版）》
CAS
2001年第1期109-112,共4页
Journal of Hebei Normal University：Natural Science
基金
全军医药卫生科学基金课题!(98Q0 76 )
