title: High level expression of a synthetic gene coding for IgG-binding domain B of Staphylococcal protein A.
abstract: A gene coding for one of the IgG-binding domains of Staphylococcal protein A, designated domain B, was chemically synthesized. This gene was tandemly repeated to give dimeric and tetrameric domain B genes by the use of two restriction enzymes which gave blunt ends. The genes were highly expressed in Escherichia coli to afford a large amount of dimeric and tetrameric domain B proteins. The single domain B protein was efficiently produced as a fusion protein with a salmon growth hormone fragment. The fusion protein was converted to monomeric domain B by cyanogen bromide cleavage. The CD spectra of the monomeric, dimeric and tetrameric domain B proteins were essentially the same as that of native form protein A, showing that their secondary structures were very similar. The dimeric and tetrameric domain B proteins formed precipitates with IgG as protein A. This system permits the efficient production of mutated single and multiple IgG-binding domains which can be used to study structural changes and protein A-immunoglobulin interactions.
authors.affiliation: Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., Tokyo, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Saito
authors.forename: S
authors.initials: S
authors.lastname: Honda
authors.forename: T
authors.initials: T
authors.lastname: Nishi
authors.forename: M
authors.initials: M
authors.lastname: Koike
authors.forename: K
authors.initials: K
authors.lastname: Okazaki
authors.forename: S
authors.initials: S
authors.lastname: Itoh
authors.forename: M
authors.initials: M
authors.lastname: Sato
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation; D005813:Genes, Synthetic; D007074:Immunoglobulin G / Q000235:genetics*; D008969:Molecular Sequence Data; D015202:Protein Engineering; D013205:Staphylococcal Protein A / Q000235:genetics*
chemical_list: D007074:Immunoglobulin G; D013205:Staphylococcal Protein A