Nucleic Acids Research, 2003, Vol. 31, No. 21 6306-6320
© 2003 Oxford University Press
Transcription profiles of the bacterium Mycoplasma pneumoniae grown at different temperatures
Zentrum für Molekulare Biologie Heidelberg, Universität Heidelberg, 69120 Heidelberg, Germany
*To whom correspondence should be addressed. Tel: +49 6221 54 68 27; Fax: +49 6221 54 58 93; Email: r.herrmann{at}mail.zmbh.uni-heidelberg.de
Present address:
H. W. H. Göhlmann, Johnson & Johnson Pharmaceutical Research and Development, 2340 Beerse, Belgium
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors
Applying microarray technology, we have investigated the transcriptome of the small bacterium Mycoplasma pneumoniae grown at three different temperature conditions: 32, 37 and 32°C followed by a heat shock for 15 min at 43°C, before isolating the RNA. From 688 proposed open-reading frames, 676 were investigated and 564 were found to be expressed (P < 0.001; 606 with P < 0.01) and at least 33 (P < 0.001; 77 at P < 0.01) regulated. By quantitative real-time PCR of selected mRNA species, the expression data could be linked to absolute molecule numbers. We found M.pneumoniae to be regulated at the transcriptional level. Forty-seven genes were found to be significantly up-regulated after heat shock (P < 0.01). Among those were the conserved heat shock genes like dnaK, lonA and clpB, but also several genes coding for ribosomal proteins and 10 genes of unassigned functions. In addition, 30 genes were found to be down-regulated under the applied heat shock conditions. Further more, we have compared different methods of cDNA synthesis (random hexamer versus gene-specific primers, different RNA concentrations) and various normalization strategies of the raw microarray data.
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