磺胺二甲氧嘧啶免疫小鼠抗体重链可变区基因的克隆及序列分析

应用RT-PCR方法从磺胺二甲氧嘧啶免疫小鼠B细胞扩增和克隆抗SDM抗体VH基因。测序结果表明:抗SDM抗体VH基因为339 bp,编码113个氨基酸。用NCBI GenBank分析表明,抗SDM抗体VH基因符合小鼠抗体重链可变区特征,隶属于小鼠抗体重链SM7家族,由胚系基因VHSM7.a3.93-DSP2.9-JH1功能性重排形成。抗SDM抗体VH基因的克隆为抗SDM单链抗体的构建与表达奠定了基础。

Cloning and Sequencing of the Heavy-Chain Variable Region's Gene Fragment from Mouse Immunized with Sulfadimethoxine

The VH gene was amplified and cloned by RT-PCR from B cell of the mouse immunized with sulfadimethoxine(SDM).The results of sequence determination showed that the VH region of antibody against SDM consisted of 339 bp encoding 113 amino acid resides.The sequence of the VH gene was homologous with those of mouse antibody variable region published in NCBI GenBank. According Kabat classified method,the VH gene of antibody against SDM belongs to the mouse Ig heavy chain subgroup SM7 and was constructed by the rearrangement of VHSM7.a3.93-DSP2.9-JH1.The success of cloning of the VH gene of antibody against SDM laid a benefit basis for the construction and expression of single-chain antibody against SDM.