Optimization of expression conditions of recombinant Fuantai-03 and detection of its biological activities / 生物医学工程学杂志
Journal of Biomedical Engineering
;
(6): 923-928, 2012.
Article
in Chinese
| WPRIM
| ID: wpr-246530
ABSTRACT
Fuantai-03(FAT-03), isolated from the Dasyatis akajei, has a strong antiangiogenic activity. The recombinant Fuantai-03 (GST/rFAT-03) fusion protein can be obtained with the DNA recombination technology. In this study, expression conditions of GST/rFAT-03 were optimized by response surface experimental design method. The constructed engineering bacteria containing GST/rFAT-03 plasmid was induced by isopropy-beta-D-thiogalactosid (IPTG), the GST affinity column was used for isolation and purification, and then the effects of different culture time, IPTG concentration, induction temperature and induction time on the amount of soluble GST/rFAT-03 fusion protein were compared. The culture time for optimal expression was 6.13 h, IPTG concentration was 0.36 mmol/L, induction temperature was 19.71 degrees C, and induction time was 13.60 h. The amount of soluble GST/rFAT-03 fusion protein was 7.57 mg/L under above mentioned expression conditions. The results also showed that rFAT-03 significantly inhibited angiogenesis in chicken chorioallantoic membrane in a dose-dependent manner. Moreover, the soluble form of the target protein is useful for further work on purification and on studying its biological function.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Recombinant Fusion Proteins
/
Skates, Fish
/
Chickens
/
Angiogenesis Inhibitors
/
Fish Proteins
/
Chorioallantoic Membrane
/
Escherichia coli
/
Genetics
/
Metabolism
Type of study:
Diagnostic study
Limits:
Animals
Language:
Chinese
Journal:
Journal of Biomedical Engineering
Year:
2012
Type:
Article
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