抗脂肪细胞特异膜蛋白单链抗体基因的克隆及序列分析  

Cloning and sequence analysis of scFv gene

作　　者：汪磊[1] 高士争[1] 张曦[1] 郝军元[2] 张玉静[2] 

机构地区：[1]云南农业大学云南省动物营养与饲料重点实验室,云南昆明650201 [2]中国人民解放军军需大学生化教研室,吉林长春130062

出　　处：《中国兽医科技》2004年第2期11-16,共6页Chinese Journal of Veterinary Science and Technology

基　　金：国家自然科学基金项目 (30 2 6 0 0 79)

摘　　要：从分泌抗 4 0ku脂肪细胞特异膜蛋白单克隆抗体的杂交瘤细胞中提取纯化mRNA ,经RT PCR扩增抗体轻、重链可变区基因 ,然后与Linker连接组装成单链抗体基因 (scFv)。结果 ,成功地构建了scFv基因 ,其排列方式为VH linker VL。经序列分析表明 ,scFv基因全长 74 1bp ,VH基因全长 36 9bp ,VL基因全长 32 7bp。With the purified mRNA from hybridoma which secrete monoclonal antibodies against a 40 ku adipocyte plasma membrane protein, the V_(H) (heavy-chain variable region) and V_(L) (light-chain variable region) genes encoding the monoclonal antibody were amplified by RT-PCR, then they were joined with Linker for construction of scFv gene. The results showed that the scFv gene had been constructed in the way of V_(H)-linker-V_(L). Sequence analysis demonstrated that length of the scFv gene was 741 bp and length of V_(H) and V_(L) genes was 369 bp and 327 bp respectively. The constructed scFv gene can be used to express the antibody.

关 键 词：脂肪细胞膜蛋白 单链抗体 序列 单克隆抗体 

分 类 号：Q26[生物学—细胞生物学]