Research Support, Non-U.S. Gov't
- Evaluation of the Cell Growth of Mycobacteria Using Mycobacterium smegmatis mc2 155 as a Representative Species
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Jorge A. Gonzalez-y-Merchand , Ruben Zaragoza-Contreras , Rosalina Guadarrama-Medina , Addy C. Helguera-Repetto , Sandra Rivera-Gutierrez , Jorge F. Cerna-Cortes , Leopoldo Santos-Argumedo , Robert A. Cox
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J. Microbiol. 2012;50(3):419-425. Published online June 30, 2012
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DOI: https://doi.org/10.1007/s12275-012-1556-0
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Abstract
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The study of the in vitro cell growth of mycobacteria still remains a fastidious, difficult, and time-consuming procedure. In addition, assessing mycobacterial growth in the laboratory is often complicated by cell aggregation and slow growth-rate. We now report that the use of a stainless steel spring in the culture led to an absence of large cell clumps, to a decrease of dead cells in the exponential phase and to growth of a more homogeneous population of large cells. We also report that flow cytometry is a rapid, simple and reliable approach to monitor mycobacterial cell growth and viability. Here, we monitored Mycobacterium smegmatis cellular growth by optical density, dry cell mass, and colony forming units; in addition, viability, cell size and granularity profiles were analyzed by flow cytometry, and cell morphology by electron microscopy. Cultures monitored by flow cytometry may lead to a better understanding of the physiology of mycobacteria. Moreover, this methodology may aid in characterizing the cell growth of other fastidious species of microorganisms.
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Citations
Citations to this article as recorded by

- Novel Populations of Mycobacterium smegmatis Under Hypoxia and Starvation: Some Insights on Cell Viability and Morphological Changes
Ruben Zaragoza-Contreras, Diana A. Aguilar-Ayala, Lázaro García-Morales, Miguel A. Ares, Addy Cecilia Helguera-Repetto, Jorge Francisco Cerna-Cortés, Lizbel León-Solis, Fernando Suárez-Sánchez, Jorge A. González-Y-Merchand, Sandra Rivera-Gutiérrez
Microorganisms.2024; 12(11): 2280. CrossRef - Screening of Hydrophilic Polymers Reveals Broad Activity in Protecting Phages during Cryopreservation
Huba L. Marton, Apoorva Bhatt, Antonia P. Sagona, Peter Kilbride, Matthew I. Gibson
Biomacromolecules.2024; 25(1): 413. CrossRef - Flow cytometry method for absolute counting and single-cell phenotyping of mycobacteria
David A. Barr, Charles Omollo, Mandy Mason, Anastasia Koch, Robert J. Wilkinson, David G. Lalloo, Graeme Meintjes, Valerie Mizrahi, Digby F. Warner, Gerry Davies
Scientific Reports.2021;[Epub] CrossRef - A Low Cost/Low Power Open Source Sensor System for Automated Tuberculosis Drug Susceptibility Testing
Kyukwang Kim, Hyeong Kim, Hwijoon Lim, Hyun Myung
Sensors.2016; 16(6): 942. CrossRef - Size evolution in microorganisms masks trade-offs predicted by the growth rate hypothesis
Isabelle Gounand, Tanguy Daufresne, Dominique Gravel, Corinne Bouvier, Thierry Bouvier, Marine Combe, Claire Gougat-Barbera, Franck Poly, Clara Torres-Barceló, Nicolas Mouquet
Proceedings of the Royal Society B: Biological Sciences.2016; 283(1845): 20162272. CrossRef - Global Adaptation to a Lipid Environment Triggers the Dormancy-Related Phenotype of Mycobacterium tuberculosis
Juan G. Rodríguez, Adriana C. Hernández, Cecilia Helguera-Repetto, Diana Aguilar Ayala, Rosalina Guadarrama-Medina, Juan M. Anzóla, Jose R. Bustos, María M. Zambrano, Jorge González-y-Merchand, María J. García, Patricia Del Portillo, Carol A. Nacy
mBio.2014;[Epub] CrossRef
- Reversible function of RapA with the C-terminus of RapC in Dictyostelium
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Dongju Kim , Wonbum Kim , Taeck Joong Jeon
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J. Microbiol. 2021;59(9):853-848.
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DOI: https://doi.org/10.1007/s12275-021-1400-5
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Abstract
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Rap small GTPases are involved in diverse signaling pathways
associated with cell growth, proliferation, and cell migration.
There are three Rap proteins in Dictyostelium, RapA, RapB,
and RapC. RapA is a key regulator in the control of cell adhesion
and migration. Recently RapA and RapC have been
reported to have opposite functions in the regulation of cellular
processes. In this study, we demonstrate that the C-terminus
of RapC, which is not found in RapA, is essential for
the opposite functions of RapC and is able to reverse the functions
of RapA when fused to the tail of RapA. Cells lacking
RapC displayed several defective phenotypes, including spread
morphology, strong adhesion, and decreased cell migration
compared to wild-type cells. These phenotypes were rescued
by full-length RapC, but not by RapC missing the C-terminus.
Furthermore, recombinant RapA fused with the C-terminus
of RapC completely recovered the phenotypes of rapC
null cells, indicating that the functions of RapA were modified
to become similar to those of RapC by the C-terminus of
RapC with respect to cell morphology, cell adhesion and migration,
cytokinesis, and development. These results suggest
that the C-terminal residues of RapC are able to suppress and
change the functions of other Ras proteins in Ras oncogenic
signaling pathways.
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Citations
Citations to this article as recorded by

- RapB Regulates Cell Adhesion and Migration in Dictyostelium, Similar to RapA
Uri Han, Nara Han, Byeonggyu Park, Taeck Joong Jeon
Journal of Microbiology.2024; 62(8): 627. CrossRef - Adhesion of Dictyostelium Amoebae to Surfaces: A Brief History of Attachments
Lucija Mijanović, Igor Weber
Frontiers in Cell and Developmental Biology.2022;[Epub] CrossRef