1887

Use of the Sucrose Gradient Method for Bacterial Cell Cycle Synchronization

    Authors: Lin Lin1, Abha Choudhary1, Anish Bavishi1, Norma Ogbonna1, Sarah Maddux1, Madhusudan Choudhary1,*
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    Affiliations: 1: Department of Biological Sciences, Sam Houston State University, Huntsville, Texas 77341
    AUTHOR AND ARTICLE INFORMATION AUTHOR AND ARTICLE INFORMATION
    • Published 03 May 2012
    • *Corresponding author. Mailing address: Department of Biological Sciences, Sam Houston State University, Avenue I, Lee Drain Building, Suite 300, P.O. Box 2116, Huntsville, Texas 77341. Phone: 936-294-4850. Fax: 936-294-3940. E-mail: mchoudhary@shsu.edu.
    • Copyright © 2012 American Society for Microbiology
    Source: J. Microbiol. Biol. Educ. May 2012 vol. 13 no. 1 50-53. doi:10.1128/jmbe.v13i1.346
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    Abstract:

    Although many undergraduate and graduate Cell and Molecular Biology courses study the bacterial cell cycle and the mechanisms that regulate prokaryotic cell division, few laboratory projects exist for the enhanced study of cell cycle characteristics in a standard teaching laboratory. One notable reason for this lack of engaging laboratory projects is, although bacterial cells can be grown fairly easily, these cultured cells are in a variety of cell cycle states. As such, to study and understand the factors that regulate bacterial cell division in morphological, physiological, and even molecular respects, it is necessary to have bacterial cells in the same stage of its cell cycle. This matching can be performed by a procedure called cell cycle synchronization.

Key Concept Ranking

Scanning Electron Microscopy
0.46891212
Bacterial Cell Division
0.4550528
0.46891212

References & Citations

1. Buknik Z, Kadlec P, Urban D, Bruhns M 1995 Sugar technologists manual: chemical and physical data for sugar manufacturers and users Bartens Publishing Co. Berlin
2. Carl PL 1970 Escherichia coli mutants with temperature-sensitive synthesis of DNA Mol. Gen. Genet. 109 107 122 10.1007/BF00269647 4925091 http://dx.doi.org/10.1007/BF00269647
3. Dwek RD, Kobrin LH, Grossman N, Ron EZ 1980 Synchronization of cell division in microorganisms by percoll gradient J. Bacteriol. 144 17 21 6252189
4. Eroglu E, Melis A 2009 “Density equilibrium” method for the quantitative and rapid in situ determination of lipid, hydrocarbon, or biopolymer content in microorganisms Biotechnol. Bioeng. 102 1406 1415 10.1002/bit.22182 http://dx.doi.org/10.1002/bit.22182
5. Evinger M, Agabian N 1977 Envelope-associated nucleoid from Caulobacter crescentus stalked and swarmer cells J. Bacteriol. 132 294 301 334726
6. Helmstetter CE, Cummings DJ 1963 Bacterial synchronization by selection of cells at division Proc. Natl. Acad. Sci. USA. 50 767 774 10.1073/pnas.50.4.767 14077509 http://dx.doi.org/10.1073/pnas.50.4.767
7. Helmstetter CE, Eenhuis C, Theisen P, Grimwade J, Leonard AC 1992 Improved bacterial baby machine: application to Escherichia coli K-12 J. Bacteriol. 174 3445 3449 1592802
8. Hesse WR, Kim MJ 2009 Visualization of flagellar interactions on bacterial carpets J. Microsc. 233 302 308 10.1111/j.1365-2818.2009.03119.x 19220696 http://dx.doi.org/10.1111/j.1365-2818.2009.03119.x
9. Kahng LS, Shapiro L 2001 The CcrM DNA methyltransferase of Agrobacterium tumefaciens is essential, and its activity is cell cycle regulated J. Bacteriol. 183 3065 3075 10.1128/JB.183.10.3065-3075.2001 11325934 http://dx.doi.org/10.1128/JB.183.10.3065-3075.2001
10. Poole RK 1977 Fluctuations in buoyant density during the cell cycle of Escherichia coli K12: significance for the preparation of synchronous cultures by age selection J. Microbiol. 98 177 186 10.1099/00221287-98-1-177 http://dx.doi.org/10.1099/00221287-98-1-177
11. Sistrom WR 1960 A requirement for sodium in the growth of Rhodopseudomonas sphaeroides J. Gen. Appl. Microbiol. 22 778 785
12. Skarstad K, Steen HB, Boye E 1983 Cell cycle parameters of slowly growing Escherichia coli B/r studied by flow cytometry J. Bacteriol. 154 656 662 6341358
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/content/journal/jmbe/10.1128/jmbe.v13i1.346
2012-05-03
2017-05-01

Abstract:

Although many undergraduate and graduate Cell and Molecular Biology courses study the bacterial cell cycle and the mechanisms that regulate prokaryotic cell division, few laboratory projects exist for the enhanced study of cell cycle characteristics in a standard teaching laboratory. One notable reason for this lack of engaging laboratory projects is, although bacterial cells can be grown fairly easily, these cultured cells are in a variety of cell cycle states. As such, to study and understand the factors that regulate bacterial cell division in morphological, physiological, and even molecular respects, it is necessary to have bacterial cells in the same stage of its cell cycle. This matching can be performed by a procedure called cell cycle synchronization.

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Figures

Image of FIGURE 1

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FIGURE 1

Buoyant densities of R. sphaeroides under different growth conditions. The determination of buoyant densities of R. sphaeroides cells both aerobically (left panel) and photosynthetically (right panel).

Source: J. Microbiol. Biol. Educ. May 2012 vol. 13 no. 1 50-53. doi:10.1128/jmbe.v13i1.346
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Image of FIGURE 2

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FIGURE 2

cells separated into two different sucrose gradient layers with corresponding buoyant densities.

Source: J. Microbiol. Biol. Educ. May 2012 vol. 13 no. 1 50-53. doi:10.1128/jmbe.v13i1.346
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Image of FIGURE 3

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FIGURE 3

Microscopic analysis of cells collected from different gradient layers and synchronized pellets: (A) cells with majority of dividing cells were collected from 68% density layers; (B) cells with evenly mixed cell populations were taken from 69% layers; (C) cells were collected from bottom pellets with highest buoyant densities. Note: Most cells were newborn.

Source: J. Microbiol. Biol. Educ. May 2012 vol. 13 no. 1 50-53. doi:10.1128/jmbe.v13i1.346
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