期刊信息
- 刊名： 河北师范大学学报（自然科学版）Journal of Hebei Normal University (Natural Science)
- 主办： 河北师范大学
- ISSN： 1000-5854
- CN： 13-1061/N
- 中国科技核心期刊
- 中国期刊方阵入选期刊
- 中国高校优秀科技期刊
- 华北优秀期刊
- 河北省优秀科技期刊
Sequence Analysis of ScFv Gene for Anti-alpha-toxin
摘要/Abstract
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2E3和1A8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1A8-ScFv,并将其克隆至pGEM-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-ScFv基因全长为729bp,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.2E3-ScFv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1A8-ScFv的VH和VL基因分别属于鼠免疫球蛋白重链(A)和轻链κ家簇
The VHand VL genes were amplified from two hybridoma cell lines producing mouse McAb against alpha-txoin of Clostridium perf ringens type A by RT-PCR.T he VHand VL genes were connected through a flexible Linker( Gly 4Ser)3,and the VH- Linker-VL(ScFv) fusion gene was cloned into a clone vector pGEM-T. The 2E3 -Sc Fv and 1A8-Sc Fv gene were sequenced and analyzed by computer.T he Sc Fv genes consist of 726 bp and 729 bp respectively.BothVH and VLgenes were functionally rearranged mouse immunoglobulin variable region genes and appeared to be new genes. According to Kabat classed method, the VH gene segment and VL gene segment of 2E3-ScFv belong to the mouse Ig heavy chain subgroupⅡ(B) and J chain subgroup Ⅲrespectively,but theVHgene segment and Lgene segment of 1A8 -ScFv belong to the mouse Ig heavy chain subgroupⅡ(A) and Jchain subgroup Ⅵ respectively.