This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow An erratum has been published
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Vandecasteele, S. J.
Right arrow Articles by Van Eldere, J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Vandecasteele, S. J.
Right arrow Articles by Van Eldere, J.

 Previous Article  |  Next Article 

Journal of Bacteriology, December 2001, p. 7094-7101, Vol. 183, No. 24
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.24.7094-7101.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Quantification of Expression of Staphylococcus epidermidis Housekeeping Genes with Taqman Quantitative PCR during In Vitro Growth and under Different Conditions

S. J. Vandecasteele,1,2,* W. E. Peetermans,1,2 R. Merckx,1 and J. Van Eldere1,3

Infectious Diseases Research Group, Department of Microbiology and Immunology, Rega Institute for Medical Research, University of Leuven,1 and Departments of General Internal Medicine2 and Microbiology and Immunology,3 University Hospital Leuven, Leuven, Belgium

Received 13 June 2001/Accepted 11 September 2001

The aims of the present study were (i) to develop and test a sensitive and reproducible method for the study of gene expression in staphylococci and (ii) to study the expression of five housekeeping genes which are involved in nucleic acid metabolism (gmk, guanylate kinase; the dihydrofolate reductase [DHFR] gene), glucose metabolism (tpi, triosephosphate isomerase), and protein metabolism (the 16S rRNA gene; hsp-60, heat-shock protein 60) during in vitro exponential and stationary growth. A modified method for instant mRNA isolation was combined with gene quantification via Taqman real-time quantitative PCR. The detection limit of our method was 10 copies of RNA. The average intersample variability was 16%. A 10-fold increase in the expression of the hsp-60 gene was induced by exposure to a 10°C heat shock (37 to 47°C) for 10 min. During in vitro growth, the expression of all five housekeeping genes showed rapid up-regulation after inoculation of the bacteria in brain heart infusion medum and started to decline during the mid-exponential-growth phase. Maximal gene expression was 110- to 300-fold higher than gene expression during stationary phase. This indicates that housekeeping metabolism is a very dynamic process that is extremely capable of adapting to different growth conditions. Expression of the 16S rRNA gene decreases significantly earlier than that of other housekeeping genes. This confirms earlier findings for Escherichia coli that a decline in bacterial ribosomal content (measured by 16S rRNA gene expression) precedes the decline in protein synthesis (measured by mRNA expression).

* Corresponding author. Mailing address: Department of General Internal Medicine, University Hospital of Leuven, Herestraat 49, B-3000 Leuven, Belgium. Phone: 32 16 34 62 74. Fax: 32 16 34 62 75. E-mail: stefaan.vandecasteele{at}uz.kuleuven.ac.be.

Journal of Bacteriology, December 2001, p. 7094-7101, Vol. 183, No. 24
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.24.7094-7101.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Shahrooei, M., Hira, V., Stijlemans, B., Merckx, R., Hermans, P. W. M., Van Eldere, J. (2009). Inhibition of Staphylococcus epidermidis Biofilm Formation by Rabbit Polyclonal Antibodies against the SesC Protein. Infect. Immun. 77: 3670-3678 [Abstract] [Full Text]  
  • Saleh-Lakha, S., Shannon, K. E., Goyer, C., Trevors, J. T., Zebarth, B. J., Burton, D. L. (2008). Nitric Oxide Reductase Gene Expression and Nitrous Oxide Production in Nitrate-Grown Pseudomonas mandelii. Appl. Environ. Microbiol. 74: 6876-6879 [Abstract] [Full Text]  
  • Ding, Y., Onodera, Y., Lee, J. C., Hooper, D. C. (2008). NorB, an Efflux Pump in Staphylococcus aureus Strain MW2, Contributes to Bacterial Fitness in Abscesses. J. Bacteriol. 190: 7123-7129 [Abstract] [Full Text]  
  • Toes, A.-C. M., Daleke, M. H., Kuenen, J. G., Muyzer, G. (2008). Expression of copA and cusA in Shewanella during copper stress. Microbiology 154: 2709-2718 [Abstract] [Full Text]  
  • Pintens, V., Massonet, C., Merckx, R., Vandecasteele, S., Peetermans, W. E., Knobloch, J. K.-M., Van Eldere, J. (2008). The role of {sigma}B in persistence of Staphylococcus epidermidis foreign body infection. Microbiology 154: 2827-2836 [Abstract] [Full Text]  
  • Zhang, H., Herman, J. P., Bolton, H. Jr., Zhang, Z., Clark, S., Xun, L. (2007). Evidence that Bacterial ABC-Type Transporter Imports Free EDTA for Metabolism. J. Bacteriol. 189: 7991-7997 [Abstract] [Full Text]  
  • Knapp, C. W., Fowle, D. A., Kulczycki, E., Roberts, J. A., Graham, D. W. (2007). From the Cover: Methane monooxygenase gene expression mediated by methanobactin in the presence of mineral copper sources. Proc. Natl. Acad. Sci. USA 104: 12040-12045 [Abstract] [Full Text]  
  • Kieboom, J., Abee, T. (2006). Arginine-Dependent Acid Resistance in Salmonella enterica Serovar Typhimurium.. J. Bacteriol. 188: 5650-5653 [Abstract] [Full Text]  
  • Sholto-Douglas-Vernon, C., Sandy, J., Victor, T. C, Sim, E., Helden, P. D. (2005). Mutational and expression analysis of tbnat and its response to isoniazid. J Med Microbiol 54: 1189-1197 [Abstract] [Full Text]  
  • Johnson, D. R., Lee, P. K. H., Holmes, V. F., Alvarez-Cohen, L. (2005). An Internal Reference Technique for Accurately Quantifying Specific mRNAs by Real-Time PCR with Application to the tceA Reductive Dehalogenase Gene. Appl. Environ. Microbiol. 71: 3866-3871 [Abstract] [Full Text]  
  • Nielsen, K. K., Boye, M. (2005). Real-Time Quantitative Reverse Transcription-PCR Analysis of Expression Stability of Actinobacillus pleuropneumoniae Housekeeping Genes during In Vitro Growth under Iron-Depleted Conditions. Appl. Environ. Microbiol. 71: 2949-2954 [Abstract] [Full Text]  
  • Richardson, J., Craighead, J. C., Cao, S. L., Handfield, M. (2005). Concurrence between the gene expression pattern of Actinobacillus actinomycetemcomitans in localized aggressive periodontitis and in human epithelial cells. J Med Microbiol 54: 497-504 [Abstract] [Full Text]  
  • Sue, D., Fink, D., Wiedmann, M., Boor, K. J. (2004). {sigma}B-dependent gene induction and expression in Listeria monocytogenes during osmotic and acid stress conditions simulating the intestinal environment. Microbiology 150: 3843-3855 [Abstract] [Full Text]  
  • Fey, A., Eichler, S., Flavier, S., Christen, R., Hofle, M. G., Guzman, C. A. (2004). Establishment of a Real-Time PCR-Based Approach for Accurate Quantification of Bacterial RNA Targets in Water, Using Salmonella as a Model Organism. Appl. Environ. Microbiol. 70: 3618-3623 [Abstract] [Full Text]  
  • Wosten, M. M. S. M., Wagenaar, J. A., van Putten, J. P. M. (2004). The FlgS/FlgR Two-component Signal Transduction System Regulates the fla Regulon in Campylobacter jejuni. J. Biol. Chem. 279: 16214-16222 [Abstract] [Full Text]  
  • Vandecasteele, S. J., Peetermans, W. E., Carbonez, A., Van Eldere, J. (2004). Metabolic Activity of Staphylococcus epidermidis Is High during Initial and Low during Late Experimental Foreign-Body Infection. J. Bacteriol. 186: 2236-2239 [Abstract] [Full Text]  
  • Savli, H., Karadenizli, A., Kolayli, F., Gundes, S., Ozbek, U., Vahaboglu, H. (2003). Expression stability of six housekeeping genes: a proposal for resistance gene quantification studies of Pseudomonas aeruginosa by real-time quantitative RT-PCR. J Med Microbiol 52: 403-408 [Abstract] [Full Text]  
  • Vandecasteele, S. J., Frans, J., Van Ranst, M., Millar, B. C., Jiru, X., Moore, J. E. (2002). Contamination Management of Broad-Range Ribosomal DNA PCR: Where Is the Evidence?. J. Clin. Microbiol. 40: 3885-3886 [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»