title: Purification and characterization of the DNA-binding protein Ner of bacteriophage Mu.
abstract: The construction is described of a plasmid (pL-ner) which directs the high-level production of the bacteriophage Mu Ner protein in Escherichia coli. The protein, recovered in the soluble cellular fraction, was susceptible to in vivo proteolytic processing, in many host strains, but not in E. coli B, a natural lon- prototroph. A simple purification method is described which takes advantage of the basic nature of the protein. The purified protein was shown to be physically and chemically homogeneous and to have an amino acid sequence identical to that predicted for the authentic protein. The protein was also shown to have in vitro biological activity, as measured by specific binding to a DNA fragment containing the consensus Ner-binding sequence, and in vivo biological activity as the protein produced by the pL-ner plasmid allowed lysogenic-like maintenance of a Mu prophage c mutant unable to synthesise a functional Mu repressor.
authors.affiliation: Biogen S.A., Geneva, Switzerland.
authors.forename: B
authors.initials: B
authors.lastname: Allet
authors.forename: M
authors.initials: M
authors.lastname: Payton
authors.forename: R J
authors.initials: RJ
authors.lastname: Mattaliano
authors.forename: A M
authors.initials: AM
authors.lastname: Gronenborn
authors.forename: G M
authors.initials: GM
authors.lastname: Clore
authors.forename: P T
authors.initials: PT
authors.lastname: Wingfield
mesh_terms: D000595:Amino Acid Sequence; D010583:Bacteriophage mu / Q000235:genetics*; D001483:Base Sequence; D004274:DNA, Recombinant; D004268:DNA-Binding Proteins / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics; D008242:Lysogeny; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014764:Viral Proteins / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D004274:DNA, Recombinant; D004268:DNA-Binding Proteins; D014764:Viral Proteins