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Expression of the synthetic gene of an artificial DDT-binding polypeptide in Escherichia coli

Moser, Rudolf ; Frey, Stefan ; Münger, Karl ; Hehlgans, Thomas ; Klauser, Stephan ; Langen, Hanno ; Winnacker, Ernst-L ; Mertz, Ronald ; Gutte, Bernd

In: Protein Engineering, Design and Selection, 1987, vol. 1, no. 4, p. 339-343

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    Titre
    • Expression of the synthetic gene of an artificial DDT-binding polypeptide in Escherichia coli
    Auteur
    • Moser, Rudolf. Biochemisches Institut der Universitāt Zūrich, Winterthurerstrasse 190, CH-8057 Zārich, Switzerland, Laboratorium für Molekulare Biologie (Genzentrum), D-8033 Martinsried, FRG
    • Frey, Stefan. Biochemisches Institut der Universitāt Zūrich, Winterthurerstrasse 190, CH-8057 Zārich, Switzerland, Laboratorium für Molekulare Biologie (Genzentrum), D-8033 Martinsried, FRG
    • Münger, Karl. Biochemisches Institut der Universitāt Zūrich, Winterthurerstrasse 190, CH-8057 Zārich, Switzerland, Laboratorium für Molekulare Biologie (Genzentrum), D-8033 Martinsried, FRG
    • Hehlgans, Thomas. Biochemisches Institut der Universitāt Zūrich, Winterthurerstrasse 190, CH-8057 Zārich, Switzerland, Laboratorium für Molekulare Biologie (Genzentrum), D-8033 Martinsried, FRG
    • Klauser, Stephan. Biochemisches Institut der Universitāt Zūrich, Winterthurerstrasse 190, CH-8057 Zārich, Switzerland, Laboratorium für Molekulare Biologie (Genzentrum), D-8033 Martinsried, FRG
    • Langen, Hanno. Biochemisches Institut der Universitāt Zūrich, Winterthurerstrasse 190, CH-8057 Zārich, Switzerland, Laboratorium für Molekulare Biologie (Genzentrum), D-8033 Martinsried, FRG
    • Winnacker, Ernst-L. Laboratorium für Molekulare Biologie (Genzentrum), D-8033 Martinsried, FRG
    • Mertz, Ronald. Biochemisches Institut der Universitāt Zūrich, Winterthurerstrasse 190, CH-8057 Zārich, Switzerland, Laboratorium für Molekulare Biologie (Genzentrum), D-8033 Martinsried, FRG
    • Gutte, Bernd. Biochemisches Institut der Universitāt Zūrich, Winterthurerstrasse 190, CH-8057 Zārich, Switzerland, Laboratorium für Molekulare Biologie (Genzentrum), D-8033 Martinsried, FRG
    Type de document
    • Postprint
    Langue
    • Anglais
    Publié dans
    • Protein Engineering, Design and Selection, 1987, vol. 1, no. 4, p. 339-343. Oxford University Press
    Autre version électronique
    Identifiant OAI-PMH
    • oai:doc.rero.ch:297107
    Summary
    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-β-D-thiogalactopyranoside a fusion protein was expressed in which DBP was linked to the COOH-termiuus of β-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 β-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