Expression and Purification of a hbFGF Lacking Nuclear Localization Signal
- VernacularTitle:一种缺失核定位信号区的hbFGF的表达及纯化
- Author:
Xiaoping WU
;
Zhijian SU
;
Qing ZHENG
;
Sixian WU
;
Ya FENG
;
Hongyan QU
;
Hua XU
;
Xiaokun LI
- Publication Type:Journal Article
- Keywords:
Human basic fibroblast growth factor;
Nuclear localization signal;
Purification
- From:
Journal of China Pharmaceutical University
2005;(3):272-275
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To study the mechanism of the unique export of one of human basic fibroblast growth factor (hbFGF) forms lacking the N-terminal nuclear localization signal (NLS),we high expressed and purified this hbFGF form in E.coli strain BL21(DE3).METHODS:The cDNA fragment of the hbFGF amplified by polymerase chain reaction (PCR) was cloned into the expression vector pET3c and expressed in BL21(DE3) by IPTG induction.The expressed hbFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate.The mitogenic activity was measured by MTT.RESULTS:The expression level of hbFGF in E.coli was about 20% of the total cellular protein.The appreciable mitogenic activity of the purified hbFGF was comparable to that of hbFGF standard.CONCLUSION:The BL21(DE3)/ pET3c expression system could be used to high express hbFGF lacking NLS.The purified recombinant hbFGF was prepared and sufficient for further study.
