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Research Support, Non-U.S. Gov't
Evaluation of the Cell Growth of Mycobacteria Using Mycobacterium smegmatis mc2 155 as a Representative Species
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
J. Microbiol. 2012;50(3):419-425.   Published online June 30, 2012
DOI: https://doi.org/10.1007/s12275-012-1556-0
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AbstractAbstract
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.

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
Dongju Kim , Wonbum Kim , Taeck Joong Jeon
J. Microbiol. 2021;59(9):853-848.
DOI: https://doi.org/10.1007/s12275-021-1400-5
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AbstractAbstract
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.

Citations

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  • 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

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