title: Fusion expression of bovine lactoferricin in Escherichia coli.
abstract: The drug resistance problem has been growing with the utilization of current antibiotics in feed and medical industries. LfcinB, a 25-amino acid antibacterial peptide derived from bovine lactoferrin, is one of potential alternatives of antibiotics. According to the bias of codon utilization of Escherichia coli, a fragment encoding LfcinB has been chemically synthesized, inserted into vector pGEX-4T-2 and expressed in E. coli. The antibacterial peptide was fused with GST with a protease cleavage site located between them. Two constructs with different cleavage sites were made. One construct, pGEX-Th-LfcinB, contains a thrombin cleavage site carried by the vector, and the other, pGEX-Th-Xa-LfcinB, contains a Factor Xa cleavage site which was introduced after the thrombin cleavage site. Fusion protein GST-Th-LfcinB protein was efficiently cleaved by thrombin, yielding recombinant LfcinB showing antibacterial activity. However, fusion protein GEX-Th-Xa-Lfcin B containing Factor Xa recognition site could not be cleaved by Factor Xa at the conditions tried in this study. Successful expression of LfcinB in E. coli provides a possible method to produce LfcinB in large amounts.
authors.affiliation: Gene Engineering Laboratory, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, PR China.
authors.forename: Xing-jun
authors.initials: XJ
authors.lastname: Feng
authors.forename: Jian-hua
authors.initials: JH
authors.lastname: Wang
authors.forename: An-shan
authors.initials: AS
authors.lastname: Shan
authors.forename: Da
authors.initials: D
authors.lastname: Teng
authors.forename: Ya-lin
authors.initials: YL
authors.lastname: Yang
authors.forename: Yi
authors.initials: Y
authors.lastname: Yao
authors.forename: Guan-pin
authors.initials: GP
authors.lastname: Yang
authors.forename: Yan-chun
authors.initials: YC
authors.lastname: Shao
authors.forename: Shuo
authors.initials: S
authors.lastname: Liu
authors.forename: Fan
authors.initials: F
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000096:biosynthesis / Q000138:chemical synthesis*; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular / Q000379:methods; D024881:Drug Resistance, Bacterial; D004926:Escherichia coli / Q000235:genetics*; D007781:Lactoferrin / Q000096:biosynthesis* / Q000138:chemical synthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000096:biosynthesis / Q000138:chemical synthesis* / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000138:chemical synthesis / Q000235:genetics*
chemical_list: D000900:Anti-Bacterial Agents; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C082171:lactoferricin B; D007781:Lactoferrin