Transcriptome of Treponema pallidum: gene expression profile during experimental rabbit infection
| Autoři | |
|---|---|
| Rok publikování | 2003 |
| Druh | Článek v odborném periodiku |
| Časopis / Zdroj | FEMS Congress of European Microbiologists |
| Fakulta / Pracoviště MU | |
| Citace | |
| Obor | Genetika a molekulární biologie |
| Klíčová slova | transcriptome; Treponema pallidum; gene expression profiling |
| Popis | DNA microarray technology was utilized to study gene expression by the syphilis spirochete Treponema pallidum subsp. pallidum (Nichols) during infection of rabbits. Microarrays containing all 1039 annotated ORFs of Treponema pallidum subspecies pallidum (Nichols) were printed on glass slides. For 1034 ORFs (out of 1039), signals higher than the threshold (average of negative control spots + 3 SDs) were detected for both RNA and DNA probes. Total RNA from T. pallidum isolated from rabbit testes 10 days post infection was labeled and standardized by cohybridization of the same arrays with treponemal chromosomal DNA labeled with a different fluorescent marker. This internal standardization technique proved to be highly reproducible and to decrease the impact of variables such as host nucleic acid contamination or variable target DNA lengths. The most highly transcribed genes were found to correlate with the most conspicuous spots identified by two dimensional gel electrophoresis, indicating that the transcript levels generally corresponded to the relative protein concentrations. Genes with high transcript concentrations included those encoding flagellar filament and cytoplasmic filament proteins, prominent lipoproteins and membrane proteins, chaperonins, proteins involved in red-ox balance, chemotaxis regulatory proteins, a V-ATPase operon, and certain metabolic enzymes such as glycolytic pathway enzymes. Independent quantitation of the expression of 84 T. pallidum genes using real-time RT-PCR approach yielded a high degree of correlation (r = 0.94). Characterization of the T. pallidum transcriptome during experimental infection provides further insight into the importance of gene expression levels in the survival and pathogenesis of this bacterium in the mammalian host. |
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