Construction and expression of anti-glioma single-chain Fv/human tumor necrosis factor α fusion gene
10.3321/j.issn:1007-8738.2001.04.027
- VernacularTitle:抗胶质瘤单链抗体－人肿瘤坏死因子α融合基因的构建及表达
- Author:
Jun DONG
;
Qiang HUANG
;
Qing LAN
;
Liying ZHOU
;
Aidong WANG
- Publication Type:Journal Article
- From:
Journal of Cellular and Molecular Immunology
2001;17(4):374-376
- CountryChina
- Language:Chinese
-
Abstract:
Aim To construct and express the fused gene of anti-glioma single chain Fv(39ScFv)/ human tumor necrosis factor α (TNF-α ). Methods The mature peptide cDNA of TNF-α was fused to 3′ -terminus of ScFv gene. The 39ScFv-TNFα fusion gene was cloned into expression vector pET20b(＋ ) and induced to express in E.coli. Results The fusion protein expressed in E.coli account for 20％ of the total bacterial protein. It displayed a single band of Mr 44 000 by reducing SDS-PAGE and Western blot, and retained both of its anti-glioma immunoreactivity and biological activity of TNF. Conclusion The fusion gene was constructed and expressed successfully in E.coli. The fusion protein possessed bifunctional activities, it may prove useful in targeting immunotherapy against glioma.
