Strong Expression of Recombinant Human Morphogenetic Protein-4 in Escherichia coli and its Bioassay in vivo
- VernacularTitle:大肠杆菌中高效表达rhBMP-4的探讨及初步活性测定
- Author:
Si-Hong GAO
;
Ju WANG
;
Que-Wei DONG
;
Kan LIU
;
Xue-Ting LIU
;
An HONG
;
Qiu-Ling XIE
;
Fen-Yong SUN
;
- Publication Type:Journal Article
- Keywords:
rhBMP-4 Site mutation Inclusion body Renaturation Activity
- From:
China Biotechnology
2006;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To produce rhBMP-4 with bioactivity in E.coli. Methods: The full-length human BMP-4 gene was mutated by PCR without changes in amino acid sequence, then the synthesized gene was cloned into plasmid pET-3c, transducted into BL21(DE)plysS, and induced by adding IPTG to a final concentration of 1.0 mmol/L. The protein product was purified using ion-exchange chromatography method and then renaturated, bioactivity was checked by C2C12 differentiation in vitro and mouse ectopic bone formation in vivo. Results: A 438 bp gene fragment encoding mature peptide of hBMP-4 was cloned , the protein product was mostly in the form of inclusion body, after renaturation, the engineering protein shows better bioactivity. Conclusion:The mutant strategy can enhance the expression of bioactive rhBMP-4 in E.coli expression system.
