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Infection and Immunity, June 2002, p. 3080-3084, Vol. 70, No. 6
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.6.3080-3084.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Construction and Phenotypic Characterization of an Auxotrophic Mutant of Mycobacterium tuberculosis Defective in L-Arginine Biosynthesis

Bhavna G. Gordhan,1 Debbie A. Smith,2 Heidi Alderton,2 Ruth A. McAdam,3 Gregory J. Bancroft,2 and Valerie Mizrahi1*

MRC/NHLS/WITS Molecular Mycobacteriology Research Unit, School of Pathology, University of the Witwatersrand and the National Health Laboratory Service, Johannesburg, South Africa and,1 Immunology Unit, London School of Hygiene & Tropical Medicine, London,2 and Respiratory Pathogens, GlaxoSmithKline Medicines Research Centre, Stevenage, United Kingdom3

Received 1 October 2001/ Returned for modification 22 November 2001/ Accepted 15 March 2002

A mutant of Mycobacterium tuberculosis defective in the metabolism of L-arginine was constructed by allelic exchange mutagenesis. The argF mutant strain required exogenous L-arginine for growth in vitro, and in the presence of 0.96 mM L-arginine, it achieved a growth rate and cell density in stationary phase comparable to those of the wild type. The mutant strain was also able to grow in the presence of high concentrations of argininosuccinate, but its auxotrophic phenotype could not be rescued by L-citrulline, suggesting that the {Delta}argF::hyg mutation exerted a polar effect on the downstream argG gene but not on argH. The mutant strain displayed reduced virulence in immunodeficient SCID mice and was highly attenuated in immunocompetent DBA/2 mice, suggesting that L-arginine availability is restricted in vivo.


* Corresponding author. Mailing address: MRC/NHLS/WITS Molecular Mycobacteriology Research Unit, NHLS (SAIMR), P.O. Box 1038, Johannesburg 2000, South Africa. Phone: 2711 4899370. Fax: 2711 4899001. E-mail: 075val{at}chiron.wits.ac.za.

Editor: S. H. E. Kaufmann


Infection and Immunity, June 2002, p. 3080-3084, Vol. 70, No. 6
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.6.3080-3084.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.




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