聚合酶链反应法扩增免疫球蛋白变区基因  

CLONING AND SEQUENCING OF IMMUNOGLOBULIN VARIABLE-REGION GENES BY POLYMERASE CHAIN REACTION

作　　者：刘健[1] 王斌[1] 潘志美[1] 田培坤[1] 张文祥[1] 

机构地区：[1]上海市肿瘤研究所免疫室,上海200032

出　　处：《江西科学》1993年第3期137-141,共5页Jiangxi Science

摘　　要：用异硫氰酸胍抽提抗人小细胞肺癌单克隆抗体杂交瘤细胞株2F7的总RNA,用逆转录酶合成第一链cDNA,用PCR技术扩增出351 bp的重链变区基因(V_H)和324bp轻链变区基因(V_L),分别克隆至pUCV_(NP)-PCR和pWR13载体上,经筛选和DNA序列分析研究,证实已获得了该单克隆抗体的变区基因.We have used acid-Guanidine Thiocyanate method to extract total RNA of hybridoma, which secreted monoclonal antibody against human small cell lung carcinoma, then synthesized first strand cDNA. We have designeda set of oligonucleotide primers to amplify the cDNA of mouse immunoglo-bulin variable-region genes by polymerase chain reaction. Here we have applied the technique to clone and sequence the variable domain of monoclonal antibody. The result shows that the lengths of light and heavy variable-region genes of this antibody are 324 and 351 base pairs respectively.

关 键 词：聚合酶链反应 免疫球蛋白 杂交瘤 

分 类 号：R730.21[医药卫生—肿瘤]