Expression, purification and biological assay of recombinant hEGF-hbFGF(78-154aa)fusion protein
- VernacularTitle:hEGF和hbFGF的C端(78-154aa)融合蛋白的表达、纯化及活性测定
- Author:
Rongjie YU
;
Ling ZHANG
;
Jian LIN
;
Qiuling XIE
;
Fenyong SUN
;
Hanlin PU
;
Zhiying LI
- Publication Type:Journal Article
- Keywords:
Human;
Epidermal growth factor-urogastrone;
Fibroblast growth factor, basic;
Recombinant fusion proteins;
Escherichia coli
- From:
Chinese Journal of Pathophysiology
1986;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To construct a recombinant hEGF-hbFGF(78-154aa)fusion protein, which not only has the heparin-binding ability, but also promotes the growth of the cells, and to express the fusion protein in E. coli expression system with high expression level.METHODS: hEGF gene was joined with 231 bp fragment coding hbFGF(78-154aa) and expressed in E. coli. The fusion protein was purified using affinity chromatography of heparin-Hyper D and analyzed with western blot. The pI value and the biological activity were both assayed.RESULTS: The fusion protein was expressed in a high expression level of about 30% of the total cell protein, as estimated by SDS-PAGE. Western analysis results showed that the antigenicity of fusion protein was similar to hEGF. Fusion protein could not only bind heparin but also promote the growth of 3T3 cell. The pI value of fusion protein was 5.2.CONCLUSION: The recombinant hEGF-hbFGF(78-154aa) fusion protein possessed the characteristics of both hEGF and hbFGF. This new-designed protein would become a good object for the research on the relationship between the structure and the function of the growth factor.
