title: Soluble expression of a synthetic gene for human translation initiation factor 4E in Escherichia coli.
abstract: In order to obtain the active form of recombinant human initiation factor (eIF) 4E effectively, an artificial synthetic gene was cloned into an expression vector (pMAL-p2) and the soluble expression was attempted in Escherichia coli under the control of a tac promoter. Two expression systems were finally constructed as a fusion protein with maltose-binding protein, which contain a recognition sequence for the site specific protease alpha-thrombin and factor Xa, respectively. Most of the fusion protein was induced as a soluble form. The soluble human eIF-4E digested from the fusion protein showed binding specificity for the m7GTP affinity column.
authors.affiliation: Department of Physical Chemistry, Osaka University of Pharmaceutical Sciences, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Morino
authors.forename: Y
authors.initials: Y
authors.lastname: Teraoka
authors.forename: M
authors.initials: M
authors.lastname: Doi
authors.forename: T
authors.initials: T
authors.lastname: Ishida
authors.forename: H
authors.initials: H
authors.lastname: Ueda
authors.forename: S
authors.initials: S
authors.lastname: Uesugi
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D039561:Eukaryotic Initiation Factor-4E; D015870:Gene Expression; D005813:Genes, Synthetic; D006801:Humans; D008969:Molecular Sequence Data; D010448:Peptide Initiation Factors / Q000235:genetics* / Q000302:isolation & purification; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification
chemical_list: D017931:DNA Primers; D039561:Eukaryotic Initiation Factor-4E; D010448:Peptide Initiation Factors; D011994:Recombinant Proteins