title: Molecular cloning, characterization and purification of ornithine carbamoyltransferase from Mycobacterium bovis BCG.
abstract: A genomic library of Mycobacterium bovis BCG has been constructed by cloning DNA partially digested with Sau3A into the Escherichia coli expression vector pAS1. The gene coding for ornithine carbamoyl-transferase (EC.2.1.3.3; OTCase), hereafter referred to as argF, was isolated from the library by complementation of a double argF-argI mutant of E. coli and its sequence was determined. The translation initiation codon used, GTG, was identified by comparing the amino acid sequence deduced from the gene with the N-terminal sequence of the corresponding purified protein. On this basis, the M. bovis BCG OTCase monomer consists of 307 amino acid residues and displays about 44% identity with other OTCases, the most closely related homologue being the anabolic enzyme of Pseudomonas aeruginosa. The native enzyme has an estimated molecular mass of 110 kDa, suggesting a trimeric structure as is the case for most of the anabolic OTCases known from various organisms.
authors.affiliation: Service de Génétique Appliquée, Université Libre de Bruxelles, Nivelles Belgium.
authors.forename: J
authors.initials: J
authors.lastname: Timm
authors.forename: I
authors.initials: I
authors.lastname: Van Rompaey
authors.forename: C
authors.initials: C
authors.lastname: Tricot
authors.forename: M
authors.initials: M
authors.lastname: Massaer
authors.forename: F
authors.initials: F
authors.lastname: Haeseleer
authors.forename: A
authors.initials: A
authors.lastname: Fauconnier
authors.forename: V
authors.initials: V
authors.lastname: Stalon
authors.forename: A
authors.initials: A
authors.lastname: Bollen
authors.forename: P
authors.initials: P
authors.lastname: Jacobs
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D009163:Mycobacterium bovis / Q000235:genetics*; D009954:Ornithine Carbamoyltransferase / Q000235:genetics* / Q000302:isolation & purification; D011401:Promoter Regions, Genetic; D016415:Sequence Alignment
chemical_list: D001426:Bacterial Proteins; D009954:Ornithine Carbamoyltransferase