Analysis of gene structure, cloning and expression of cyp51 from Ustilago maydis / 生物工程学报
Chinese Journal of Biotechnology
; (12): 1747-1753, 2008.
Article
in Chinese
| WPRIM (Western Pacific)
| ID: wpr-275345
Responsible library:
WPRO
ABSTRACT
The cyp51 primers and two pairs of mutant primers which removed different transmembrane region were designed based on Ustilago maydis cyp51 gene structure analysis. The full cyp51 DNA fragment as well as mutant cyp51 genes were amplified and cloned by using the total DNA from Ustilago maydis as template, then subcloned into different expression vectors. The recombinant expression plasmids were transformed into Escherichia coli BL21 (DE3), BL21 (DE3) pLysS and Rosetta (DE3) respectively. A series of experiments leads to the finding that only pET32-YH-35 could be highly expressed at the optimal condition of 30 degrees C induced with 0.5 mmol/L IPTG The expressed protein (CYP51) showed biological activity by spectra analysis of the protein binding to 4 standard fungicides and to 14 XF-synthetic fungicide compounds, and only one XF-synthetic fungicide compound (XF-113) was similar to standard fungicides in binding constant. This compound is promising to be a new effective antifungal drug. These results will facilitate the further study on the mechanism of pathogenic fungi CYP51 and pesticide molecules, and will provide a new idea for efficient design and development of new anti-fungal drugs.
Full text:
Available
Health context:
Neglected Diseases
Health problem:
Neglected Diseases
/
Zoonoses
Database:
WPRIM (Western Pacific)
Main subject:
Plasmids
/
Ustilago
/
Recombinant Proteins
/
Fungal Proteins
/
Cloning, Molecular
/
Cytochrome P-450 Enzyme System
/
Escherichia coli
/
Sterol 14-Demethylase
/
Genetics
/
Metabolism
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2008
Document type:
Article