抗口蹄疫病毒单克隆抗体1C7重轻链可变区基因的克隆及序列分析

Cloning and Sequencing of the Varibale Region's Gene Fragments of the Anti-FMDV Monoclonal Antibody 1C7

应用分子生物学技术 ,从分泌抗O型口蹄疫病毒单克隆抗体的杂交瘤细胞 1C7中提取总RNA ,经反转录 ,PCR扩增及克隆 ,分别得到VH 基因及VL 基因。序列测定结果表明 :1C7的VH 基因为 36 8bp ,VL 基因为 32 3bp。用NCBIGenBank分析表明 ,VH 和VL 均符合小鼠抗体可变区特征 ,为功能性重排的抗体可变区基因。根据Kabat分类体系 ,1C7的VH 基因中的VH基因片段隶属于抗体重链第 7183家族 ,其VL 基因中的VL基因片段隶属于抗体K轻链 2 0家族 ,VH 基因由VH76_1BG_DFL16 .1_JH4重排而形成 ,VL 基因由VKbw2 0_JK2重排而形成。 1C7的VH 和VL

The V H and V L genes were amplified from a hybridoma cell line 1C7 producing mouse McAb against foot-and-mouth disease virus by RT-PCR.The results of sequence determination showed that PCR products of V H region consisted of 368 bp encoding about 122 amino acid resides,and PCR products of V L region contained 323 bp encoding about 107 amino acid resides.The sequences of V H and V L genes were homologous with those of mouse antibody variable region published in NCBI Genbank.According to Kabat classified method,mAb 1C7 V H and V L gene segments belong to the mouse Ig heavy chain subgroup VH 7183 and K chain subgroup V K 20 respectively.1C7 V H was constructed by the rearrangement of VH76-1BG-DFL16.1-JH4 and 1C7 V L by the rearrangement of V K bw20-JK2.The success of cloning of the gene fragments of mAb 1C7's variable regions laid a benefit basis for the construction and expression of anti-FMDV single-chain Fv.