title: [Expression of Pleurocidin from winter flounder in Escherichia coli and optimization of culture conditions].
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.
authors.forename: Xuejiao
authors.initials: X
authors.lastname: Xu
authors.forename: Xiangdong
authors.initials: X
authors.lastname: Zha
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Che
authors.forename: Lijuan
authors.initials: L
authors.lastname: Ma
authors.forename: Siqun
authors.initials: S
authors.lastname: Wu
authors.forename: Peilong
authors.initials: P
authors.lastname: Yang
authors.forename: Huoqing
authors.initials: H
authors.lastname: Huang
authors.forename: Bin
authors.initials: B
authors.lastname: Yao
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism*; D029941:Fish Proteins / Q000096:biosynthesis*; D005432:Flounder; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*
chemical_list: D029941:Fish Proteins; D011993:Recombinant Fusion Proteins; C105851:pleurocidin