建立Ig基因富集的DNA库进行2F7单克隆抗体基因克隆  

Coustruction of 2E7 DNA Warehouse with Ig Gene Enriched Conemic DNA and the Coding Gene Cloning of 2E7 monoclonal Antibody

作　　者：郭婵[1] 潘志美[1] 刘健[1] 洪锦心 

机构地区：[1]上海市肿瘤研究所免疫室,200032

出　　处：《单克隆抗体通讯》1990年第2期1-4,共4页

摘　　要：用小鼠型抗人小细胞肺癌单克隆抗体的2F7杂交瘤细胞DNA,经限制性内切酶EcoRI或BamHI完全酶解,通过Southern blot,用小鼠Ig基因Cr_(2a)、V_(H)、J_(H)和E_(H)作探针,富集阳性DNA,同时用pSV_2neo质粒为载体与这些DNA重组,经筛选获得了一批杂交阳性的克隆。In order to make easy manioulation in gonomic DNA warehouse construction, the Ig gene enriched DNA warehouse was constructed for the coding gene of 2E7 monoclonal antibody which reacted with human small cell lung carcinoma. The Ig gene enriched DNA was isolated from the positive band of mouse r2a probing Eco Rl digested 2E7 genomic DNA in Southern blot, and alternative collected from 6-12 Kb gradient ultracentrifuged BamHI cleaved 2E7 genomic DNA. Using 32P labelled mouse r2a, JH and EH (enhancer) probes, some, Ig VH genss were cloned. The PCR ( polymerase chain reaction reaction ) test with VH 3' and 5' terminal primer also demostrated that these colones seemed to be the 2E7 functional coding gene.

关 键 词：免疫球蛋白 DNA库 单克隆抗体 

分 类 号：R392.11[医药卫生—免疫学]