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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。
Total RNA was extracted from the hybridoma cells secreting anti-Cysticercus cellulosae monoclonal antibody, which was used as a template to amplify the light and heavy chain variable region genes of anti-Cysticercus cellulosae monoclonal antibody by RT-PCR. The results of agarose gel electrophoresis showed that about 350 bp, in line with the characteristics of the mouse antibody. The PCR product was transformed into JM109 after being ligated with pMD18-T, and the positive recombinant was screened by the blue-white spot method. Positive recombinants were identified using universal primers and sequenced. The results showed that the light chain variable region and heavy chain variable region genes were in accordance with the light chain variable region and the heavy chain variable region of the murine antibody. There are three CDR regions and FR regions in VH and VL.