MEM INST OSWALDO CRUZ, RIO DE JANEIRO, Vol. 112 | 2017
PAGES: DOI: 10.1590/0074-02760170105 Short communication
Construction of Mycobacterium tuberculosis cdd knockout and evaluation of invasion and growth in macrophages

Anne Drumond Villela1,+, Valnês S Rodrigues-Junior1, Antônio Frederico Michel Pinto1, Virgínia Carla de Almeida Falcão1,2, Zilpa Adriana Sánchez-Quitian1,2, Paula Eichler1, Cristiano Valim Bizarro1,2, Luiz Augusto Basso1,2,3, Diógenes Santiago Santos1,2

1Pontifícia Universidade Católica do Rio Grande do Sul, Centro de Pesquisas em Biologia Molecular e Funcional, Instituto Nacional de Ciência e Tecnologia em Tuberculose, Porto Alegre, RS, Brasil
2Pontifícia Universidade Católica do Rio Grande do Sul, Programa de Pós-Graduação em Biologia Celular e Molecular, Porto Alegre, RS, Brasil
3Pontifícia Universidade Católica do Rio Grande do Sul, Programa de Pós-Graduação em Medicina e Ciências da Saúde, Porto Alegre, RS, Brasil

Abstract

Cytidine deaminase (MtCDA), encoded by cdd gene (Rv3315c), is the only enzyme identified in nucleotide biosynthesis pathway of Mycobacterium tuberculosis that is able to recycle cytidine and deoxycytidine. An M. tuberculosis knockout strain for cdd gene was obtained by allelic replacement. Evaluation of mRNA expression validated cdd deletion and showed the absence of polar effect. MudPIT LC-MS/MS data indicated thymidine phosphorylase expression was decreased in knockout and complemented strains. The cdd disruption does not affect M. tuberculosis growth both in Middlebrook 7H9 and in RAW 264.7 cells, which indicates that cdd is not important for macrophage invasion and virulence.

Financial support: INCT-TB (Brazil), Decit/SCTIE/MS-MCT-CNPq-FNDCT-CAPES, CNPq (441720/2014-5 to ADV), BNDES (14.2.0914.1).
+ Corresponding author: This e-mail address is being protected from spambots. You need JavaScript enabled to view it.
Received 15 March 2017
Accepted 30 May 2017

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