title: Expression of the synthetic gene of an artificial DDT-binding polypeptide in Escherichia coli.
abstract: This paper reports the expression of an artificial functional polypeptide in bacteria. The gene of a designed 24-residue DDT-binding polypeptide (DBP) was inserted between the BamHI and PstI cleavage sites of plasmid pUR291. The hybrid plasmid, pUR291-DBP, was cloned in Escherichia coli JM109. After induction by isopropyl-beta-D-thiogalactopyranoside a fusion protein was expressed in which DBP was linked to the COOH-terminus of beta-galactosidase. DBP, which is stable to trypsin, was obtained by tryptic digestion of the fusion protein and subsequent fractionation of the tryptic peptides by reversed-phase h.p.l.c. Recombinant and chemically synthesized DBP showed identical chromatographic properties, amino acid composition, and chymotryptic digestion patterns. Both the beta-galactosidase-DBP fusion and isolated recombinant DBP bound DDT. The fusion protein was 25 times as potent as the designed 24-residue DBP in activating a cytochrome P-450 model system using equimolar catalytic amounts of the two proteins.
authors.affiliation: Biochemisches Institut der Universität Zürich, Switzerland.
authors.forename: R
authors.initials: R
authors.lastname: Moser
authors.forename: S
authors.initials: S
authors.lastname: Frey
authors.forename: K
authors.initials: K
authors.lastname: Münger
authors.forename: T
authors.initials: T
authors.lastname: Hehlgans
authors.forename: S
authors.initials: S
authors.lastname: Klauser
authors.forename: H
authors.initials: H
authors.lastname: Langen
authors.forename: E L
authors.initials: EL
authors.lastname: Winnacker
authors.forename: R
authors.initials: R
authors.lastname: Mertz
authors.forename: B
authors.initials: B
authors.lastname: Gutte
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002352:Carrier Proteins; D003634:DDT / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics*; D005786:Gene Expression Regulation; D005813:Genes, Synthetic; D008969:Molecular Sequence Data; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification
chemical_list: D002352:Carrier Proteins; D011993:Recombinant Fusion Proteins; D003634:DDT