title: A versatile plasmid expression vector for the production of biotinylated proteins by site-specific, enzymatic modification in Escherichia coli.
abstract: A versatile plasmid vector was designed to direct the synthesis of recombinant proteins in either one of two forms that will be biotinylated in Escherichia coli with high efficiency at a single, unique site. The protein of interest can be produced with a peptide substrate for E. coli biotin holoenzyme synthetase (BirA) joined directly to its N terminus, or alternatively, as a fusion to the C terminus of a maltose-binding protein domain (MalE) with the peptide substrate on its N terminus. To maximize the yield of biotinylated protein, the vector is designed to express the substrate in a coupled translation arrangement with the enzyme.
authors.affiliation: Roche Research Center, Nutley, NJ 07110, USA.
authors.forename: K L
authors.initials: KL
authors.lastname: Tsao
authors.forename: B
authors.initials: B
authors.lastname: DeBarbieri
authors.forename: H
authors.initials: H
authors.lastname: Michel
authors.forename: D S
authors.initials: DS
authors.lastname: Waugh
mesh_terms: D018528:ATP-Binding Cassette Transporters; D000595:Amino Acid Sequence; D001483:Base Sequence; D001710:Biotin; D002352:Carrier Proteins / Q000737:chemistry; D017931:DNA Primers / Q000737:chemistry; D004926:Escherichia coli; D029968:Escherichia coli Proteins; D005822:Genetic Vectors; D058619:Maltose-Binding Proteins; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins; D033902:Periplasmic Binding Proteins; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification*; D013466:Sulfurtransferases / Q000378:metabolism
chemical_list: D018528:ATP-Binding Cassette Transporters; D002352:Carrier Proteins; D017931:DNA Primers; D029968:Escherichia coli Proteins; C461619:MalE protein, E coli; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D033902:Periplasmic Binding Proteins; D011994:Recombinant Proteins; C461599:maltose transport system, E coli; D001710:Biotin; D013466:Sulfurtransferases; C057985:biotin synthetase