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Biology

Caractérisation biophysique des fonctions flagellaire moteur

Published: January 18, 2017 doi: 10.3791/55240

Materials

Name Company Catalog Number Comments
Poly-L-lysine Solution (0.1%) Sigma-Aldrich P8920 http://www.sigmaaldrich.com/catalog/product/sigma/p8920?lang=en&region=US
Polybead Microspheres Polysciences, Inc. 7307 http://www.sigmaaldrich.com/catalog/product/sigma/p8920?lang=en&region=US
1 mL Luer Slip Tip Syringe Exel Int. 26048 http://www.exelint.com/tuberculin_syringes.php
Clay Adams Intramedic Luer-Stub Adapter 23-gauge Becton, Dickinson and Company 427565 http://www.bd.com/ds/productCenter/ES-LuerStubAdaptors.asp
Polyethylene tubing Harvard Apparatus 59-8325 http://www.harvardapparatus.com/laboratory-polye-polyethylene-non-sterile-tubing.html
Photomultiplier Tubes Hamamatsu R7400U-20 Spectral response range of 300 to 920 nm, Peak wavelength 630 nm,  0.78 ns response time 
http://pdf1.alldatasheet.com/datasheet-pdf/view/212308/HAMAMATSU/R7400U-20.html
3 x 1 mm precision slits Edmund Optics NT39-908 2 slits mounted at right angles to one another on photomultiplier tubes
Oscilloscope Tektronix TBS 1032B Alternative brands are acceptable. Digital Oscilloscope, TBS 1000B Series, 2 Analogue, 30 MHz, 500 MSPS, 2.5 kpts 
http://www.tek.com/oscilloscope/tbs1000b-digital-storage-oscilloscope
8 Pole LP/HP Filter Krohn-Hite 3384 Alternative brands are acceptable. A frequency range from 0.1 Hz to 200 kHz is recommended.   
http://www.krohn-hite.com/htm/filters/PDF/3384Data.pdf
Optiphot microscope Nikon NA Any upright or inverted phase microscope can be used.
https://www.thorlabs.com/newgrouppage9.cfm?objectgroup_id=754
50:50 (R:T) Cube Beamsplitter ThorLabs BS013

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References

  1. Emody, L., Kerenyi, M., Nagy, G. Virulence factors of uropathogenic Echerichia coli. Int J Antimicrob. Ag. 22, 29-33 (2003).
  2. Lane, M. C., et al. Role of motility in the colonization of uropathogenic Escherichia coli in the urinary tract. Infect Immun. 73 (11), 7644-7656 (2005).
  3. Kao, C. Y., et al. The complex interplay among bacterial motility and virulence factors in different Escherichia coli infections. Eur J Clin Microbiol Infect Dis. 33 (12), 2157-2162 (2014).
  4. Berg, H. C. The rotary motor of bacterial flagella. Annu Rev Biochem. 72, 19-54 (2003).
  5. McCarter, L., Hilmen, M., Silverman, M. Flagellar Dynamometer Controls Swarmer Cell Differentiation of V. parahaemolyticus. Cell. 54 (3), 345-351 (1988).
  6. Lele, P. P., Hosu, B. G., Berg, H. C. Dynamics of mechanosensing in the bacterial flagellar motor. Proc Natl Acad Sci U S A. 110 (29), 11839-11844 (2013).
  7. Gode-Potratz, C. J., Kustusch, R. J., Breheny, P. J., Weiss, D. S., McCarter, L. L. Surface sensing in Vibrio parahaemolyticus triggers a programme of gene expression that promotes colonization and virulence. Mol Microbiol. 79 (1), 240-263 (2011).
  8. Kearns, D. B. A field guide to bacterial swarming motility. Nat Rev Microbiol. 8 (9), 634-644 (2010).
  9. Belas, R. Biofilms, flagella, and mechanosensing of surfaces by bacteria. Trends Microbiol. 22 (9), 517-527 (2014).
  10. Silverman, M., Simon, M. Flagellar rotation and the mechanism of bacterial motility. Nature. 249, 73-74 (1974).
  11. Block, S. M., Segall, J. E., Berg, H. C. Adaptation Kinetics in Bacterial Chemotaxis. J Bacteriol. 154 (1), 312-323 (1983).
  12. Segall, J. E., Block, S. M., Berg, H. C. Temporal comparisons in bacterial chemotaxis. Proc Natl Acad Sci U S A. 83, 8987-8991 (1986).
  13. Blair, D. F., Berg, H. C. Restoration of torque in defective flagellar motors. Science. 242 (4886), 1678-1681 (1988).
  14. Ryu, W. S., Berry, R. M., Berg, H. C. Torque-generating units of the flagellar motor of Escherchia coli have a high duty ratio. Nature. 403, 444-447 (2000).
  15. Yuan, J., Berg, H. C. Resurrection of the flagellar rotary motor near zero load. Proc Natl Acad Sci U S A. 105 (4), 1182-1185 (2008).
  16. Yuan, J., Fahrner, K. A., Berg, H. C. Switching of the bacterial flagellar motor near zero load. J Mol Biol. 390 (3), 394-400 (2009).
  17. Sowa, Y., Hotta, H., Homma, M., Ishijima, A. Torque-speed Relationship of the Na+-driven Flagellar Motor of Vibrio alginolyticus. J Mol Biol. 327 (5), 1043-1051 (2003).
  18. Xing, J., Bai, F., Berry, R., Oster, G. Torque-speed relationship of the bacterial flagellar motor. Proc Natl Acad Sci U S A. 103 (5), 1260-1265 (2006).
  19. Meacci, G., Tu, Y. Dynamics of the bacterial flagellar motor with multiple stators. Proc Natl Acad Sci U S A. 106 (10), 3746-3751 (2009).
  20. Lele, P. P., Roland, T., Shrivastava, A., Chen, Y. H., Berg, H. C. The flagellar motor of Caulobacter crescentus generates more torque when a cell swims backwards. Nat Phys. 12 (2), 175-178 (2016).
  21. Lele, P. P., Shrivastava, A., Roland, T., Berg, H. C. Response thresholds in bacterial chemotaxis. Sci Adv. 1 (9), e1500299 (2015).
  22. Berg, H. C., Turner, L. Torque Generated by the Flagellar Motor of Escherichia coli. Biophys J. 65, 2201-2216 (1993).
  23. Bai, F., et al. Conformational Spread as a Mechanism for Cooperativity in the Bacterial Flagellar Switch. Science. 327, 685-689 (2010).
  24. Reid, S. W., et al. The maximum number of torque-generating units in the flagellar motor of Escherichia coli is at least 11. Proc Natl Acad Sci U S A. 103, 8066-8071 (2006).
  25. Chen, X., Berg, H. C. Torque-Speed Relationship of the Flagellar Rotary Motor of Escherichia coli. Biophys J. 78, 1036-1041 (2000).
  26. Turner, L., Caplan, S. R., Berg, H. C. Temperature-induced switching of the bacterial flagellar motor. Biophys J. 71, 2227-2233 (1996).
Caractérisation biophysique des fonctions flagellaire moteur
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Cite this Article

Ford, K. M., Chawla, R., Lele, P. P. More

Ford, K. M., Chawla, R., Lele, P. P. Biophysical Characterization of Flagellar Motor Functions. J. Vis. Exp. (119), e55240, doi:10.3791/55240 (2017).

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