人黑色素瘤特异性单克隆抗体可变区基因的克隆和序列测定

Amplification, cloning and sequencing of the variable region genes of a human melanoma specific monoclonal antibody

作者从培养的可分泌人黑色素瘤特异性单克隆抗体的杂交瘤细胞Leo Mel3提取RNA,反转录成cDNA.用合成的寡核苷酸引物从中扩增了抗体的轩、重链可变区基因,并克隆入质粒.随机挑取阳性克隆各一个进行核苷酸序列分析.见所克隆基因确可编码小鼠抗体的轻、重链可变区。

Total RNA was extracted from a cultured hybridoma cell line, Leo Mel 3, which secretes human melanoma specific monoclonal antibody, and subjected to reverse transcription. Variable region genes of the antibody light and heavy chains were amplified from the cDNA with pairs of synthetic oligomers as primers, cloned, and sequenced. The data indicate that the cloned genes encode the variable regions of light and heavy chain of mouse antibody, respectively.