抗人小细胞肺癌单克隆抗体的重链变区基因的体外扩增、克隆和序列分析  

Cloning and Sequencing of Immunoglobulin Heavy Variableregion Gene by Polymerase Chain Reaction

作　　者：刘健[1] 吴文书[1] 王斌[1] 潘志美[1] 田培坤[1] 黄震宇[1] 洪锦心 

机构地区：[1]上海市肿瘤研究所免疫室,上海200032

出　　处：《生物化学杂志》1993年第1期70-75,共6页

摘　　要：本文从抗人小细胞肺癌单克隆抗体杂交瘤细胞中抽提总RNA,合成第一链cDNA,直接用PCR技术(polymerase chain reaction)扩增出351bp的重链变区基因(V_H),克隆至pUCV_(NP)-PCR载体上,经筛选得一批插入片段为351bp的阳性克隆,经核苷酸序列分析研究,证实已获得了该单克隆抗体的重链可变区基因。We have designed a set of oligonucleotide primers to amplify the cDNA of mouse immunoglobulin heavy variable-region gene by polymerase chain reaction. The primers incorporate restriction sites that allow the cDNA of the variable domain to be force-cloned for sequencing and expression. Here we have applied the technique to clone and sequence the heavy variable domain of monoclonal antibody that reacts with human small cell lung carcinoma line NCI H128. The result shows that the length of heavy variable-region gene of this antibody is 351 base pairs.

关 键 词：聚合酶链反应 重链变区 嵌合抗体 

分 类 号：R392.1[医药卫生—免疫学]