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A Two-Step Strategy for the Complementation of M. Tuberculosis Mutants

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journal contribution
posted on 06.03.2012, 00:00 by Farahnaz Movahedzadeh, Rosangela Frita, Hiten J. Gutka
The sequence of Mycobacterium tuberculosis, completed in 1998, facilitated both the development of genomic tools, and the creation of a number of mycobacterial mutants. These mutants have a wide range of phenotypes, from attenuated to hypervirulent strains. These phenotypes must be confirmed, to rule out possible secondary mutations that may arise during the generation of mutant strains. This may occur during the amplification of target genes or during the generation of the mutation, thus constructing a complementation strain, which expresses the wild-type copy of the gene in the mutant strain, becomes necessary. In this study we have introduced a two-step strategy to construct complementation strains using the Ag85 promoter. We have constitutively expressed dosR and have shown dosR expression is restored to wild-type level.


This work was funded by the Wellcome Trust grant 073237 and American Lung Association (Grant ID Number RG-82534-N).


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This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The original version is available through the Brazilian Genetics Society at DOI: 10.1590/S1415-47572011000200020.


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