title: Production and purification of recombinant human oxytocin overexpressed as a hybrid protein in Escherichia coli.
abstract: The plasmid DNA pERilox4 containing the gene of the recombinant protein, which included the leader sequence and the oxytocinoyl lysine tetramer, was constructed. The high level of gene expression in E. coli was achieved. The method for purification of the recombinant protein and its isolation in the soluble form was developed. The conditions for digestion of the hybrid protein by trypsin and carboxypeptidase B were matched. The effective method for transformation of oxytocinic acid to oxytocin was worked out. The scheme suggested allowed obtaining oxytocin in high yield.
authors.affiliation: Shemyakin Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow, 117997 Russia. esipov@ibch.ru
authors.forename: Roman S
authors.initials: RS
authors.lastname: Esipov
authors.forename: Larisa A
authors.initials: LA
authors.lastname: Chupova
authors.forename: Sergey V
authors.initials: SV
authors.lastname: Shvets
authors.forename: Dmitry V
authors.initials: DV
authors.lastname: Chuvikovsky
authors.forename: Alexandr I
authors.initials: AI
authors.lastname: Gurevich
authors.forename: Tatyana I
authors.initials: TI
authors.lastname: Muravyova
authors.forename: Anatoly I
authors.initials: AI
authors.lastname: Miroshnikov
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D006801:Humans; D007377:Interleukin-3 / Q000235:genetics; D008969:Molecular Sequence Data; D010121:Oxytocin / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D014169:Transformation, Bacterial
chemical_list: D007377:Interleukin-3; D011993:Recombinant Fusion Proteins; D010121:Oxytocin