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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 isolate the light chain variable region (VL) gene of LC9-G5, an anti-human laryngeal cancer monoclonal antibody, and analyze its nucleotide sequence. Methods The VL gene of anti-human laryngeal carcinoma monoclonal antibody LC9-G5 was amplified by PCR in vitro and cloned into PMD18-T vector. The recombinant plasmid was sequenced and the sequence was analyzed by NCBI and Kabat. Results The sequence was 99% homologous to the mouse ovarian cancer antibody VL gene. The full-length gene was 336 bp in length and belongs to the class II subclass of mouse immunoglobulin light chain. The sequence of the VL gene was included in Gene Bank No. U60463. Conclusion This gene is a LC9-G5 light chain variable region gene of anti-human laryngeal carcinoma monoclonal antibody.