title: Characterization of pNI10 plasmid in Pseudomonas, and the construction of an improved Escherichia and Pseudomonas shuttle vector, pNUK73.
abstract: The complete nucleotide sequence of pNI10 (3.75 kb), from which pNI105 and pNI107 were constructed as medium-host-range vectors for Gram-negative bacteria, was determined. A fragment of about 2.1 kb of pNI10 was essential for replication in Escherichia coli and Pseudomonas fluorescens. This fragment encodes a putative origin of replication ( ori) and one putative replication-controlling protein (Rep). An improved version of the medium-host-range plasmid vector pNUK73 (5.13 kb) was constructed with the basic-replicon of pNI10 and pHSG298 (2.68 kb). We show that expression in pseudomonads of the bromoperoxidase gene ( bpo) of Pseudomonas putida, inserted downstream of the lac promoter in pNUK73, resulted in about 30% (13.6 U/l culture) of the enzyme level obtained in E. coli.
authors.affiliation: Biotechnology Research Center, Toyama Prefectural University, Kurokawa 5180, 939-0398, Kosugi, Toyama, Japan. itoh@pu-toyama.ac.jp
authors.forename: N
authors.initials: N
authors.lastname: Itoh
authors.forename: T
authors.initials: T
authors.lastname: Kawanami
authors.forename: C
authors.initials: C
authors.lastname: Nitta
authors.forename: N
authors.initials: N
authors.lastname: Iwata
authors.forename: S
authors.initials: S
authors.lastname: Usami
authors.forename: Y
authors.initials: Y
authors.lastname: Abe
authors.forename: Y
authors.initials: Y
authors.lastname: Koide
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D004925:Escherichia / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D011549:Pseudomonas / Q000235:genetics*; D016415:Sequence Alignment
chemical_list: D001426:Bacterial Proteins