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

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368bp,VL基因为323bp。用NCBI GenBank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20家族，VH基因由VH76－1BG－DFL16．1－JH4重排而形成，VL基因由KVbw20-JK2重排而形成。1C7的VH和VL基因的克隆为抗FMDV scFv的构建与表达奠定了基础。

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

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.