High and stable expression of an analog of human basic fibroblast growth factor in Escherichia coli / 中国病理生理杂志
Chinese Journal of Pathophysiology
;
(12): 247-250, 2006.
Artigo
em Chinês
| WPRIM
| ID: wpr-408800
ABSTRACT
AIM:
To obtain a high and stable expression analog of human basic fibroblast growth factor by genetic engineering.METHODS:
The cysteins 78 and 96 of natural hbFGF polypeptide was substituted with serines by means of site-directed mutagenesis. Using pET- 3c as vector, the mutated polynucleotide was cloned and then transferred into BL21 (DE3)plysS. After induction by IPTG, the analog was obtained and analyzed by SDS - PAGE.RESULTS:
After purification the form of soluble mutant increased remarkably but the forms of dimmer and higher multimer were reduced greatly to no more than 8% of the total recombinant protein. By MTT assay, the analog showed the same biological activity. This new analog represented a desirable complementation for native hbFGF to develop pharmaceutical drug in clinical use.CONCLUSION:
Substitution of certain amino acids of polypeptide without altering native protein' s bioactivity to get the analog is an effective means to increase stability of foreign protein and its solubility in E. coli.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Idioma:
Chinês
Revista:
Chinese Journal of Pathophysiology
Ano de publicação:
2006
Tipo de documento:
Artigo
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