Expression of Pleurocidin from winter flounder in Escherichia coli and optimization of culture conditions / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 365-374, 2016.
Article
in Chinese
| WPRIM
| ID: wpr-337407
ABSTRACT
To express Pleurocidin in Escherichia coli and to enhance the secretory efficiency of the fusion protein, the gene encoding Pleurocidin was ligated with Cherry DNA sequence via blunt-end ligation. Then this fusion gene was cloned into pET22b (+) vector and the recombinant plasmid was transformed into E. coli BL21 (DE3). Lactose was used to induce expression of fusion protein. The recombinant plasmid pET22b (+) -CP was successfully constructed and high-level expression of fusion protein was induced with lactose. Statistics showed that addition of glycine after 16 h of induction significantly enhanced the secretory efficiency of the fusion protein. After hydrolysis of the fusion protein by diluted hydrochloric acid and some further purification steps, r-Pleurocidin was obtained with antibacterial activity against E. coli DH5α and Bacillus subtilis BS168. In conclusion, the fusion protein was expressed in E. coli and biologically active r-Pleurocidin was obtained after hydrochloric acid cleavage and purification.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Flounder
/
Recombinant Fusion Proteins
/
Cloning, Molecular
/
Fish Proteins
/
Escherichia coli
/
Metabolism
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2016
Type:
Article
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