Transcriptome of Treponema pallidum: gene expression profiling of treponemes grown in rabbits
| Autoři | |
|---|---|
| Rok publikování | 2003 |
| Druh | Článek ve sborníku |
| Konference | Sborník příspěvků VII. Pracovní setkání biochemiků a molekulárních biologů |
| Fakulta / Pracoviště MU | |
| Citace | |
| Obor | Genetika a molekulární biologie |
| Klíčová slova | transcriptome; Treponema pallidum; gene expression profiling |
| Popis | DNA microarray chips containing all 1039 annotated ORFs of Treponema pallidum subsp. pallidum (Nichols) were printed on glass slides. For 1034 ORFs (out of 1039), signals higher than threshold (average of negative control spots + 3 SDs) were detected for both labeled RNA and DNA. Total RNA isolated from treponemes grown in rabbit testes was labeled and standardized to the labeled treponemal chromosomal DNA. Genes were sorted according to the relative transcription level and the operon structure of T. pallidum genome was proposed. The most intensively transcribed genes were found to correlate with most prominent spots identified on 2D SDS PAGE gels indicating that the transcription rate approximately corresponds to the level of protein synthesis. Gene expression of 84 genes was independently measured using real-time PCR approach and the results were compared to the data obtained by DNA microarray technique. In addition, expression levels of treponemal genes were measured using large insert library of T. pallidum DNA in E. coli. Significant gene expression differences were found and the average level of gene expression in E. coli computed for 10-gene windows appears to correlate inversely to the number of individual gene copies in the library. These data indicate that the level of gene expression in the host organism is one of the major factors contributing to efficiency of DNA cloning. |
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