Expression and Purification of Basic Fibroblast Growth Factor Mutant with Reduced Mitogenic Activity
10.3969/j.issn.1671-8135.2005.02.010
- VernacularTitle:一种促分裂活性降低的hbFGF突变体的表达及纯化
- Author:
Xiao-Ping WU
1
;
Xiao-Kun LI
;
Zhi-Jian SU
;
Qing ZHENG
;
Si-Xian WU
;
Hua XU
;
Hong-Yan QU
Author Information
1. 暨南大学
- Keywords:
Human basic fibroblast growth factor Mutation Purification Mitogenic activity
- From:
China Biotechnology
2005;25(2):49-52
- CountryChina
- Language:Chinese
-
Abstract:
In order to decrease the potential side-effects of human basic fibroblast growth factor (hbFGF) caused by its broadspectrum mitogenic activity, a single residue of hbFGF, the residue serine 108, was replaced with neutral alanine residue to construct a mutant of hbFGF (mhbFGF) with reduced mitogenic activity. The mutant was overexpressed in Escherichia coli BL21(DE3) by IPTG induction. The expression level of mhbFGF was about 30% of the total cellular protein. The expressed mhbFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate. Measured by MTT method, the effect of mhbFGF on Balb/c 3T3 cell proliferation was much lower than that of wild-type hbFGF. The purified recombinant mhbFGF was prepared and sufficient for the following pharmacological and safety studies.
