抗HFRSV人单抗可变区基因克隆及其序列测定  

Gene Cloning and Sequencing of the Variable Region of Human Monoclonal Antibody Against Hemorrhagic Fever Virus

作　　者：高磊[1] 陈苏民[1] 陈南春[1] 杨安钢[1] 崔运昌[1] 

机构地区：[1]第四军医大学生物化学及分子生物学教研室,西安710032

出　　处：《生物工程学报》1994年第4期338-341,共4页Chinese Journal of Biotechnology

基　　金：国家自然科学基金;编号39080021

摘　　要：从杂交瘤细胞株87-2提取细胞总RNA,反转录合成cDNA链,进行PCR反应。其中扩增重链可变区一对引物分别与人免疫球蛋白重链V区基因5＇端和J区基因3＇端互补;扩增人λ轻链可变区引物与人免疫球蛋白λ轻链V区基因5＇端和J区基因3＇端互补。将重、轻链可变区基因的PCR扩增产物分别插入M13噬菌体,经转化筛选分别获得重组克隆。双脱氧法测定其序列,所得核苷酸序列经计算机分析,轻、重链可变区基因长度分别为309bp和405pb,编码103个氨基酸和135个氨基酸,有明显抗体可变区特征,具有骨架区和抗原互补区。Human-Human hybridoma cell line 87-2 secretes anti HFRSV human monoclonal antibody. Total RNA was extracted from the cell and was reverse transcribed to the first strand cDNA using oligo d (T) as primer. A set of oligqnucleotide primers were designed to amplify the cDNA of human immunoglubulin heavy and light chain variable domains by polymerase chain reaction. The amplified VH and VL fragments were ligat-ed to the phage M13 vector DNA. The clones containing V gene inserts were sequenced. The sequences were confirmed as the human immunoglubulin variable genes by comparing with those of known sequences . There are framework (FR) and complementarity determining (CDR) regions in the variable genes.

关 键 词：肾病综合征 出血热 病毒 单克隆抗体 基因工程 

分 类 号：R394.8[医药卫生—医学遗传学] R392.11[医药卫生—基础医学]