鼠源抗胃癌抗体轻链可变区基因克隆及其结构分析

Cloning and constructive analysis of mouse anti-gastric-cancer antibody light chain variable region gene fragments

目的 扩增与人胃癌细胞特异结合的鼠源性抗体轻链可变区基因 ,并对其基因序列进行分析 .方法 用 RT-PCR方法从分泌鼠源抗人胃癌抗体的杂交瘤细胞扩增出抗体轻链可变区基因 ,克隆到 p GEM- T载体 ,测序仪测定其基因序列并对其结构进行分析 .结果 所获得的基因由 35 4bp组成 ,编码 118个氨基酸 ,序列分析证实该基因可变区序列符合小鼠 Ig轻链可变区的氨基酸序列特征 .结论 获得的鼠源抗胃癌抗体轻链可变区基因为κ 家族 。

AIM To amplify and to analyze the light chain variable gene fragments (V H) of mouse antigastriccancer antibody. METHODS V H gene fragments of mouse antigastriccancer antibody were amplified by RTPCR method by using RNA purified from antibody producing hybridoma cells, and further cloned into pGEMT vector which were then sequenced. RESULTS V H gene fragment consisted of 354 bp and encoded 118 amino acid residues and the amino acid sequences of variable region was in agreement with the characteristics of the amino acid present in the mouse Ig variable region. CONCLUSION The light chain variable gene fragments of mouse antigastriccancer antibody belong to κⅢ subgroup and the successful cloning of this gene fragment might be used in the experiments of immunotherapeutic study of gastriccancer.