摘要
目的:获得抗人视网膜母细胞瘤单克隆抗体轻链可变区基因。方法:从分泌抗人视网膜母细胞瘤单克隆抗体的杂交瘤细胞中提取总RNA,逆转录形成cDNA,用扩增小鼠K轻链可变区基因的引物,通过RT-PCR方法,扩增基因,1.5%琼脂糖凝胶电泳鉴定。结果:用轻链引物扩增获得340bp的基因片段。结论:从三株杂交瘤细胞中均获得高纯度的总RNA;用RT-PCR方法从一株杂交瘤细胞成功克隆了抗人视网膜母细胞瘤单克隆抗体轻链可变区基因,为基因工程抗体研究奠定了良好基础。眼科学报2001;17:194~197。
Objective: To acquire the variable region gene of light chain of monoclonal antibodyagainst human retinoblastoma.Methods: Total RNA were extracted from hybridoma cells secreting specific monoclonalantibody (MeAb) against human retinoblastoma (RB), then transcripted reversely intocDNA with random primers. The variable region of the light chain (VL) gene fragmentswere ampliflied using polymerase chain reaction (PCR) method. Agrose gelelectrophoresis was confirmed.Results: 1.5% agrose gel electrophoresis indicated that VL. gene was about 340 basepairs.Conclusion: The light chain variable region gene of the McAb against human RB wasamplified successfully, which lays a good basis for construction of a recombinantantibody. Eye Science 2001; 17: 194 ~ 197.
出处
《眼科学报》
2001年第4期194-197,共4页
Eye Science
基金
国家自然科学基金(资助号:39870801)
广东省自然科学基金(资助号:98011)
中山医科大学"211"工程项目(资助号:98007)资助