抗猪囊尾蚴单克隆抗体轻重链可变区基因的扩增及序列分析

Amplified and sequence analysis of V_H and V_L genes of monoclonal antibody against Cysticercus cellulosae

从分泌抗猪囊尾蚴单克隆抗体杂交瘤细胞提取总RNA,以此为摸板,RT-PCR扩增出抗猪囊尾蚴单克隆抗体轻重链可变区基因,琼脂糖凝胶电泳检测显示约350 bp,符合鼠抗体特征。PCR产物与pMD18-T连接后转化JM109,蓝白斑法筛选出阳性重组子。以载体通用引物对阳性重组子进行鉴定,对证明为阳性的重组子测序。结果显示,轻链可变区和重链可变区基因符合鼠抗体轻链可变区和重链可变区基因特征,VH和VL有3个比较明显的CDR区和FR区。

The VH and VL genes were amplified from the hybridoma cell secreting monoclonal antibody against Cysticercus cellulosae by RT-PCR. The length of VH and VL was about 350 bp by agarose gel eleetrophoresis,which corresponded with the characterizations of mouse antibody. PCR products were linked with pMD18-T vector, and the cloned plasmids were transformed into JM109. The positive clones were selected by blue/white blot,then were iden tiffed by common primer. The result of sequencing showed the gene sequence accorded with the VH and VL genes characterization of mouse antibody and contained three remarkable CDR and FR.