title: Construction, expression, and characterization of recombinant hirudin in Escherichia coli.
abstract: The mutant gene of HV2-K47 was obtained by polymerase chain reaction-directed mutagenesis and expressed in Escherichia coli. Many elements that could affect its expression level were compared. The product was purified to homogeneity via three chromatographic steps--ion exchange, gel filtration, and reverse phase chromatography--on the AKTA Explorer System. The anti-thrombin activity of HV2-K47 is much higher than that of recombinant HV2. Some properties and expression conditions were investigated systematically, which would be useful for further studies of hirudin and other small proteins.
authors.affiliation: College of Life Sciences, Peking University, Beijing, PR China.
authors.forename: Q
authors.initials: Q
authors.lastname: Bi
authors.forename: J
authors.initials: J
authors.lastname: Zhang
authors.forename: Y
authors.initials: Y
authors.lastname: Huang
authors.forename: H
authors.initials: H
authors.lastname: Su
authors.forename: X
authors.initials: X
authors.lastname: Zhou
authors.forename: S
authors.initials: S
authors.lastname: Zhu
mesh_terms: D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D005343:Fibrinolytic Agents; D006629:Hirudins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D016296:Mutagenesis; D021382:Protein Sorting Signals / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
chemical_list: D005343:Fibrinolytic Agents; D006629:Hirudins; D021382:Protein Sorting Signals; D011994:Recombinant Proteins