Gene Cloning, Construction and Expression of Single-Chain Fv (scFv) Against the Membrane Protein of Schisotosoma japonicum
- VernacularTitle:抗日本血吸虫膜蛋白特异性单链抗体的构建及表达
- Author:
Xiaocong YU
;
Xin JIANG
;
Haomin HUANG
;
Zhong ZHANG
;
Qing LIN
;
Xiaohong GUAN
;
Hualiang HUANG
- Publication Type:Journal Article
- Keywords:
single-chain antibody, variable region genes, construction, expression, Schistosoma japonicum
- From:
Chinese Journal of Parasitology and Parasitic Diseases
1987;0(03):-
- CountryChina
- Language:Chinese
-
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.
