1887

Abstract

Transcripts of the gas vesicle genes and were detected in single filaments of the cyanobacterium using reverse transcription and quantitative real-time PCR. Primers were designed to amplify short sequences within and three length variants of . With genomic template DNA, and using Sybr Green to monitor product accumulation, similar amplification efficiencies were observed for each of these genes. The relative copy numbers of length variants in genomic DNA from five gas vesicle genotypes determined by real-time PCR were similar to those indicated by sequencing the gas vesicle gene clusters. The precipitation of cDNA reverse-transcribed from cellular RNA from single filaments was required before amplification of the gene fragments; without this step it was not possible to detect the accumulation of the expected amplicons by dissociation analysis. Precipitation was also necessary to ensure the generation of product curves that allowed linear regression in an early stage of PCR, a prerequisite for the quantification of low-input cDNA amounts without the need for standard curves. This report shows that different length variants are transcribed within single filaments, both from laboratory cultures and from natural samples taken from Lake Zürich. This has implications for the efficiency of buoyancy provision by the possible production of gas vesicles of different strengths within individual cyanobacterial filaments. The hypothesis that post-transcriptional regulation may influence the type of protein (GvpC) present in gas vesicles is presented.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.27402-0
2005-01-01
2024-12-07
Loading full text...

Full text loading...

/deliver/fulltext/micro/151/1/mic1510059.html?itemId=/content/journal/micro/10.1099/mic.0.27402-0&mimeType=html&fmt=ahah

References

  1. Albouy D., Castets A. M., Tandeau De Marsac N. 2001; The gas vesicle gene (gvp) cluster of the cyanobacterium Pseudanabaena sp. strain PCC 6901. DNA Seq 12:337–344
    [Google Scholar]
  2. Beard S. J., Handley B. A., Hayes P. K., Walsby A. E. 1999; The diversity of gas vesicle genes in Planktothrix rubescens from Lake Zürich. Microbiology 145:2757–2768
    [Google Scholar]
  3. Beard S. J., Davis P. A., Walsby A. E, Iglesias-Rodríguez D., Skulberg O. M. 2000; Gas vesicle genes in Planktothrix spp. from Nordic lakes: strains with weak gas vesicles possess a longer variant of gvpC . Microbiology 146:2009–2018
    [Google Scholar]
  4. Becker S., Ernst A, Böger P., Oehlmann R. 2000; PCR bias in ecological analysis: a case study for quantitative Taq nuclease assays in analyses of microbial communities. Appl Environ Microbiol 66:4945–4953 [CrossRef]
    [Google Scholar]
  5. Bright D. I., Walsby A. E. 1999; The relationship between critical pressure and width of gas vesicles in isolates of Planktothrix rubescens from Lake Zürich. Microbiology 145:2769–2775
    [Google Scholar]
  6. Bright D. I., Walsby A. E. 2000; The daily integral of growth by Planktothrix rubescens calculated from growth rate in culture and irradiance in Lake Zürich. New Phytol 146:301–316 [CrossRef]
    [Google Scholar]
  7. Damerval T., Houmard J., Guglielmi G., Csiszàr K., Tandeau de Marsac N. 1987; A developmentally regulated gvpABC operon is involved in the formation of gas vesicles in the cyanobacterium Calothrix 7601. Gene 54:83–92 [CrossRef]
    [Google Scholar]
  8. Griffiths A. E., Walsby A. E., Hayes P. K. 1992; The homologies of gas vesicle proteins. J Gen Microbiol 138:1243–1250 [CrossRef]
    [Google Scholar]
  9. Hayes P. K., Powell R. S. 1995; The gvpA/C cluster of Anabaena flos-aquae has multiple copies of a gene encoding GvpA. Arch Microbiol 164:50–57 [CrossRef]
    [Google Scholar]
  10. Heid C. A., Stevens J., Livak K. J., Williams P. M. 1996; Real-time quantitative PCR. Genome Res 6:986–994 [CrossRef]
    [Google Scholar]
  11. Klein D., Janda P., Steinborn R., Müller M., Salmons B., Günzburg W. H. 1999; Proviral load determination of different feline immunodeficiency virus isolates using real-time polymerase chain reaction: influence of mismatches on quantification. Electrophoresis 20:291–299 [CrossRef]
    [Google Scholar]
  12. Kromkamp J., Walsby A. E. 1990; A computer model of buoyancy and vertical migration in cyanobacteria. J Plankton Res 12:161–183 [CrossRef]
    [Google Scholar]
  13. Liss B. 2002; Improved quantitative real-time RT-PCR for expression profiling of individual cells. Nucleic Acids Res 30:e89 [CrossRef]
    [Google Scholar]
  14. Livak K. J., Schmittgen T. D. 2001; Analysis of relative gene expression data using real-time quantitative PCR and the [inline-graphic] method. Methods 25:402–408 [/inline-graphic] [CrossRef]
    [Google Scholar]
  15. Oliver R. L. 1994; Floating and sinking in gas-vacuolate cyanobacteria. J Phycol 30:161–173 [CrossRef]
    [Google Scholar]
  16. Oliver R. L., Walsby A. E. 1984; Direct evidence for the role of light-mediated gas vesicle collapse in the buoyancy regulation of Anabaena flos-aquae (cyanobacteria). Limnol Oceanogr 29:879–886 [CrossRef]
    [Google Scholar]
  17. Pfeifer F., Gregor D., Hofacker A., Plosser P., Zimmermann P. 2002; Regulation of gas vesicle formation in halophilic archaea. J Mol Microbiol Biotechnol 4:175–181
    [Google Scholar]
  18. Ramakers C., Ruijter J. M., Deprez R. H., Moorman A. F. 2003; Assumption-free analysis of quantitative real-time polymerase chain reaction (PCR) data. Neurosci Lett 339:62–66 [CrossRef]
    [Google Scholar]
  19. Shukla H. D., DasSarma S. 2004; Complexity of gas vesicle biogenesis in Halobacterium sp. strain NRC-1: identification of five new proteins. J Bacteriol 186:3182–3186 [CrossRef]
    [Google Scholar]
  20. Skulberg O. M., Skulberg R. 1985; Planktic species of Oscillatoria (Cyanophyceae) from Norway. Characterization and classification. Arch Hydrobiol Suppl 71:157–174
    [Google Scholar]
  21. Utkilen H. C., Oliver R. L., Walsby A. E. 1985; Buoyancy regulation in a red Oscillatoria unable to collapse gas vacuoles by turgor pressure. Arch Hydrobiol 102:319–329
    [Google Scholar]
  22. Walsby A. E. 1994; Gas vesicles. Microbiol Rev 58:94–144
    [Google Scholar]
  23. Walsby A. E., Avery A. 1996; Measurement of filamentous cyanobacteria by image analysis. J Microbiol Methods 26:11–20 [CrossRef]
    [Google Scholar]
  24. Walsby A. E., Bleything A. 1988; The dimensions of cyanobacterial gas vesicles in relation to their efficiency in providing buoyancy and withstanding pressure. J Gen Microbiol 134:2635–2645
    [Google Scholar]
  25. Walsby A. E., Avery A., Schanz F. 1998; The critical pressures of gas vesicles in Planktothrix rubescens in relation to the depth of winter mixing in Lake Zürich. Switzerland. J Plankton Res 20:1357–1375 [CrossRef]
    [Google Scholar]
/content/journal/micro/10.1099/mic.0.27402-0
Loading
/content/journal/micro/10.1099/mic.0.27402-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error