›› 2001, Vol. 19 ›› Issue (3): 2-140.

• 论著 • Previous Articles     Next Articles

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

YU Xiao-cong1;JIANG Xin1;HUANG Hao-min1;ZHANG Zhong1;LIN Qing1;GUAN Xiao-hong2;HUANG Hua-liang1*   

  1. 1 Institute of Genetics;Chinese Academy of Sciences;Beijing 100101; 2 Department of Parasitology;Nanjing Medical University;N
  • Received:1900-01-01 Revised:1900-01-01 Online:2001-06-30 Published:2001-06-30

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Abstract: 　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.

Key words: single-chain antibody, variable region genes, construction, expression, Schistosoma japonicum