Strong Expression of Recombinant Human Morphogenetic Protein-4 in Escherichia coli and its Bioassay in vivo / 中国生物工程杂志
China Biotechnology
;
(12)2006.
Article
in Chinese
| WPRIM
| ID: wpr-684884
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
China Biotechnology
Year:
2006
Type:
Article
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