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Molecular cloning, characterization and purification of ornithine carbamoyltransferase from Mycobacterium bovis BCG

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Summary

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 carbamoyltransferase (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.

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Author notes

  1. Juliano Timm

    Present address: Unité de Génie Microbiologique, Institut Pasteur, 28 rue du Docteur Roux, F-7015, Paris, France

Authors and Affiliations

  1. Service de Génétique Appliquée, Université Libre de Bruxelles, rue de l'Industrie 24, B-1400, Nivelles, Belgium

    Juliano Timm, Ingrid Van Rompaey, Marc Massaer, Françoise Haeseleer, Alan Fauconnier, Alex Bollen & Paul Jacobs

  2. Université Libre de Bruxelles et Institut de recherches du Centre d'Enseignement et de Recherches des Industries Alimentaires et Chimiques, Av. E. Gryzon 1, B-1070, Brussels, Belgium

    Catherine Tricot & Victor Stalon

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  1. Juliano Timm
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  2. Ingrid Van Rompaey
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  3. Catherine Tricot
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  4. Marc Massaer
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  9. Paul Jacobs
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Timm, J., Van Rompaey, I., Tricot, C. et al. Molecular cloning, characterization and purification of ornithine carbamoyltransferase from Mycobacterium bovis BCG. Molec. Gen. Genet. 234, 475–480 (1992). https://doi.org/10.1007/BF00538708

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