Biochemical and Biophysical Research Communications
Functional characterization of Mycobacterium tuberculosis Rv2969c membrane protein
Section snippets
Materials and methods
Bioinformatics analysis. Mycobacterium tuberculosis Rv2969c protein sequences were obtained from the Tuberculist Webserver (http://www.genolist.pasteur.fr/TubercuList/). Protein alignments were done with BLAST and protein features and GRAVY scores were calculated by using the PROTPARAM tool (http://www.us.expasy.org/tools/protparam.html) [10]. Signal sequences, non-classical secretion, transmembrane domains, and O-glycosylation sites were predicted
Mycobacterial species and strains. The ATCC and
Molecular analysis of the Rv2969c gene
The presence of Rv2969c in MTC (M. tuberculosis H37Rv, M. tuberculosis H37Ra, M. bovis, M. bovis BCG, M. africanum, and M. microti) strains was confirmed by the amplification of a 587 bp region from this gene (Fig. 1A). A similar amplification band was also seen in 10 M. tuberculosis isolates from patients suffering pulmonary and extra-pulmonary tuberculosis (data not shown). Sequence analysis of all clinical isolates PCR amplicons revealed no sequence variation suggesting that this protein was
Acknowledgments
This research has been supported by COLCIENCIAS contract RC-2008. Nora Martinez translated this manuscript.
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2011, PeptidesCitation Excerpt :It has been reported that a considerable number of secreted proteins are protective antigens, and therefore have been considered as attractive candidates to develop subunit vaccines [3,33,36], or proteins that bind to the host cell and that can be used in designing diagnostic methods [62] Moreover, they are hypothesized to mediate mycobacterial entry into the host cell [59]. The methodology used here for identifying binding fragments from the selected proteins has been previously applied in P. falciparum studies [4,28,30,31,44,55,68,69], Plasmodium vivax [48,67] and M. tuberculosis [12,18,26,49,50,53,58,70]. In the case of P. falciparum, identified sequences have been the base to the rational design of subunit-based, multiantigenic, multistage chemically synthesized vaccines, and a universal methodology has been proposed, that might be applied to prevent different infectious diseases [13,51,52].
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