抗人喉癌单克隆抗体轻链可变区基因的克隆

Cloning of the light chain variable region gene of monoclonal antibody against human laryngeal carcinoma

目的分离抗人喉癌单克隆抗体LC9-G5轻链可变区(light chain variable region,VL)基因,并测定分析其核苷酸序列。方法使用PCR方法体外扩增抗人喉癌单克隆抗体LC9-G5 VL基因,将其克隆入PMD18-T载体,重组子测序,将其序列使用数据库NCBI和Kabat进行分析。结果这段序列与小鼠卵巢癌抗体VL基因的同源性为99%,基因全长336 bp,属于小鼠免疫球蛋白轻链第Ⅱ亚类,该VL基因序列已被Gene Bank收录,收录号为U60463。结论该基因为抗人喉癌单克隆抗体LC9-G5轻链可变区基因。

OBJECTIVE To amplify and sequence the light chain variable region（VL） gene of monoclonal antibody （McAb） against human laryngeal carcinoma. METHODS The VL gene of mouse McAb against human laryngeal carcinoma was amplified by polymerase chain reaction（PCR）. The PCR product was then cloned into PMD18-T vector. The recombinants were sequenced. The VL gene was compared with published mouse VL gene by NCBI and Kabat. RESULTS It was proved that a full-length VL gene was 336 bp, the VL gene was a member of mouse immunoglobulin （Ig）light chain subgroup Ⅱ. The VL gene sequence was registered by Gene Bank （accession No. U60463）. CONCLUSION The obtained VL gene is potentially the functional gene of the McAb against human laryngeal carcinoma.