单克隆抗体可变区基因PCR扩增产物的直接测序

利用一组通用引物对单克隆抗体可变区ｃＤＮＡ进行ＰＣＲ扩增。扩增产物经纯化后通过一个以链终止法为基础的“循环测序”程序直接进行序列测定，其中所用四种双脱氧核苷酸分别带不同的荧光标记，测序引物选自同一组通用引物。测序电泳和数据收集在ＤＮＡ测序仪上进行。本法简单、快速、准确，是解析抗体可变区序列的有效手段。

Direct Sequencing of PCR-amplified Monoclonal Antibody Variable Region Genes

Monoclonal antibody variable region cDNAs were amplified through PCR using a group of universal variable region primers. Products of amplification were directly sequenced by chain termination-based cycle sequencing protocol after being purified, in which four ddNTPs were separately labeled with four different fluoresceins,and primers used for PCR amplification were reemployed as sequencing primers.Sequencing electrophoresis and data collection were performed on DNA sequencer.Being simple, time-saving and reliable, this approach is powerful in the dissection ofantibody variable region sequences.