title: Expression of a synthetic gene coding for the amino acid sequence of Clostridium pasteurianum rubredoxin.
abstract: A synthetic gene based on the published amino acid sequence for Clostridium pasteurianum rubredoxin was constructed, cloned in Escherichia coli 71/18 and expressed using the T7 RNA polymerase/promoter system in E. coli HMS273. UV/visible spectroscopy and metal analyses indicated that the as-isolated synthetic gene product is a mixture of holo-(i.e. iron-containing) rubredoxin and zinc-substituted rubredoxin, with the latter amounting to approximately 70% of the total rubredoxin. The UV/visible absorption and resonance Raman spectra of the cloned holorubredoxin are characteristic of the native rubredoxin-type iron site. N-terminal amino acid sequencing suggests that the gene product consists of at least three polypeptide species with the initial sequences (approximate relative abundances): Met-Met-Lys-... (63%), blocked (30%) and Met-Lys-... (7%). The blocked portion presumably consists of a mixture of nMet-Met-Lys-... and nMet-Lys-..., where nMet represents an amino-blocked methionine residue.
authors.affiliation: Department of Chemistry, University of Georgia, Athens 30602.
authors.forename: M K
authors.initials: MK
authors.lastname: Eidsness
authors.forename: S E
authors.initials: SE
authors.lastname: O'Dell
authors.forename: D M
authors.initials: DM
authors.lastname: Kurtz
authors.forename: R L
authors.initials: RL
authors.lastname: Robson
authors.forename: R A
authors.initials: RA
authors.lastname: Scott
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001483:Base Sequence; D003013:Clostridium / Q000235:genetics; D005813:Genes, Synthetic; D008969:Molecular Sequence Data; D015202:Protein Engineering; D012416:Rubredoxins / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D001426:Bacterial Proteins; D012416:Rubredoxins; C077196:rubredoxin protein, Clostridium pasteurianum