High and stable expression of an analog of human basic fibroblast growth factor in Escherichia coli
- VernacularTitle:重组人碱性成纤维细胞生长因子结构类似物在大肠杆菌中的高效稳定表达研究
- Author:
Qiongyu CHEN
;
Fenyong SUN
;
Xiaojia CHEN
;
Qiuling XIE
;
Jinhua SUN
;
An HONG
- Publication Type:Journal Article
- Keywords:
Fibroblast growth factor 2;
Construction analog;
Escherichia coli
- From:
Chinese Journal of Pathophysiology
2006;22(2):247-250
- CountryChina
- Language:Chinese
-
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.
