抗日本血吸虫膜蛋白特异性单链抗体的构建及表达  

Gene Cloning, Construction and Expression of Single-Chain Fv (scFv) Against the Membrane Protein of Schisotosoma japonicum

作　　者：俞小淙[1] 蒋欣[1] 黄浩旻 张众[1] 林晴[1] 管晓虹[2] 黄华樑[1] 

机构地区：[1]中国科学院遗传研究所,北京100101 [2]南京医科大学寄生虫学教研组,南京210029

出　　处：《中国寄生虫学与寄生虫病杂志》2001年第3期135-140,共6页Chinese Journal of Parasitology and Parasitic Diseases

摘　　要：目的　用基因工程抗体技术构建抗日本血吸虫膜蛋白特异的单链抗体。　方法　用抗体框架区的通用引物 PCR扩增抗日本血吸虫膜蛋白特异性单克隆抗体 NP11- 4的 VH 及 VL 基因 ,测序分析其核苷酸序列。用VH 和 VL 基因在 p THA90质粒载体中构建成单链抗体基因并诱导表达。　结果　扩增获得 NP11- 4的 VH 和 VL基因 ,经测序分析确定为新发现的抗体可变区序列 ,构建成排列顺序为 VH- linker- VL 的单链抗体基因 ,经诱导表达出与硫氧环蛋白融合的单链抗体 ,大小约为 36 .2 k Da,以包涵体形式存在。　结论　成功地构建和表达了抗日本血吸虫膜蛋白特异性单链抗体。Objective To construct single chain antibody specific to membrane protein of Schistosoma japonicum by gonetic engineering technique. Methods The V\-H (heavy-chain variable region) and V\-L (light-chain variable region) genes were amplified by PCR from the genomic DNA of NP11-4 cell line, and sequenced by Sanger's method. The ScFv was constructed in pTHA90 vector using V\-H and V\-L genes, then expressed by IPTG. Results The V\-H and V\-L genes were obtained through PCR. The DNA sequences showed that V\-H and V\-L were new variable region genes of antibody. They were registered by GenBank. A ScFv gene with (Gly4Ser) 3 intralinker in the pTHA90 vector was successfully constructed. The ScFv was expressed as thioredoxin-fused proteins about 36\^2 kDa. Conclusion A specific ScFv against the membrane protein of Schistosoma japonicum was constructed and expressed.

关 键 词：单链抗体 可变区基因 日本血吸虫病 抗日本血吸虫膜蛋白特异性单链抗体 

分 类 号：R532.21[医药卫生—内科学]