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
em Zh
| WPRIM
| ID: wpr-337407
Biblioteca responsável:
WPRO
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.
Texto completo:
1
Índice:
WPRIM
Assunto principal:
Linguado
/
Proteínas Recombinantes de Fusão
/
Clonagem Molecular
/
Proteínas de Peixes
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Escherichia coli
/
Metabolismo
Limite:
Animals
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2016
Tipo de documento:
Article