Coupling reporter expression to respiration detects active as well as dormant mycobacteria in vitro and in mouse tissues

International Journal of Mycobacteriology
Subash Chand VermaManju Yasoda Krishnan

Abstract

Mycobacterium tuberculosis is known to slow down its transcriptional activity during dormancy. Hence, while using reporter strains, it is important to couple the reporter gene to a promoter that is strong and sensitive both in active and dormant M. tuberculosis. Since respiration is an indispensable process even in dormant bacteria, validation of the promoters of respiratory chain genes - type II NADH dehydrogenase (Pndh) and adenosine triphosphate (ATP) synthase operon (Patps) - of MTB was undertaken for this purpose. Putative promoter containing sequences were cloned upstream of a red fluorescent protein (RFP) gene. Mycobacterium smegmatis or M. tuberculosis carrying episomal constructs were validated for growth, fitness and fluorescence in different models in vitro and in vivo. Either promoter can drive stable and strong expression of RFP in actively growing and dormant M. smegmatis in vitro without significantly affecting growth or viability. Fluorescence due to Pndh and Patps was significantly higher than Phsp60. The fitness of M. tuberculosis H37Rv counterparts was unaffected inside J774 macrophages. In immunocompetent mice, despite an initial attenuation in the lungs, both strains reached loads similar to wild type durin...Continue Reading

References

Jul 22, 1998·FEMS Microbiology Letters·T DickB Murugasu-Oei
Dec 29, 1998·Journal of Bacteriology·M J SmeuldersH D Williams
Sep 4, 2003·The Journal of Experimental Medicine·Dirk SchnappingerGary K Schoolnik
Sep 4, 2003·The Journal of Experimental Medicine·Martin I VoskuilGary K Schoolnik
Nov 5, 2003·Nature Medicine·John D McKinney, James E Gomez
Dec 13, 2003·Tuberculosis·James E Gomez, John D McKinney
Mar 16, 2005·Proceedings of the National Academy of Sciences of the United States of America·Edward A WeinsteinHarvey Rubin
Mar 19, 2005·Journal of Bacteriology·Sougata Roy, Parthasarathi Ajitkumar
Oct 18, 2005·Proceedings of the National Academy of Sciences of the United States of America·Lanbo ShiMaria L Gennaro
Dec 22, 2006·PloS One·Olivier NeyrollesBrigitte Gicquel
Jun 19, 2007·Nature Methods·Ekaterina M MerzlyakDmitriy M Chudakov
Jul 16, 2008·The Journal of Biological Chemistry·Anil KoulKoen Andries
Aug 14, 2008·Proceedings of the National Academy of Sciences of the United States of America·Srinivasa P S RaoKevin Pethe
Dec 11, 2008·Journal of Visualized Experiments : JoVE·Renan Goude, Tanya Parish
Apr 1, 2009·Microbiology·Aleksey M AnuchinArseny S Kaprelyants
Dec 17, 2009·Tuberculosis·Ying KongJeffrey D Cirillo
May 25, 2012·Vaccine·UNKNOWN WHO Publication
May 29, 2012·The Journal of Antimicrobial Chemotherapy·Andrea ZelmerUlrich E Schaible
Aug 2, 2012·Angewandte Chemie·Daria M ShcherbakovaVladislav V Verkhusha
May 2, 2013·The Journal of Antimicrobial Chemotherapy·Nuria AndreuBrian D Robertson

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