2003
Transcriptome of Treponema pallidum: gene expression profiling of treponemes grown in rabbits
ŠMAJS, David, Petra MATĚJKOVÁ, SJ NORRIS a GM WEINSTOCKZákladní údaje
Originální název
Transcriptome of Treponema pallidum: gene expression profiling of treponemes grown in rabbits
Název česky
Transcriptom Treponema pallidum: expresní profilování během experimentální infekce králíků
Název anglicky
Transcriptome of Treponema pallidum: gene expression profiling of treponemes grown in rabbits
Autoři
ŠMAJS, David (203 Česká republika, garant), Petra MATĚJKOVÁ (203 Česká republika), SJ NORRIS (840 Spojené státy) a GM WEINSTOCK (840 Spojené státy)
Vydání
2003. vyd. Brno, Sborník příspěvků VII. Pracovní setkání biochemiků a molekulárních biologů, s. 12-12, 2003
Nakladatel
Masarykova univerzita
Další údaje
Jazyk
čeština
Typ výsledku
Stať ve sborníku
Obor
Genetika a molekulární biologie
Stát vydavatele
Česká republika
Utajení
není předmětem státního či obchodního tajemství
Kód RIV
RIV/00216224:14110/03:00012307
Organizační jednotka
Lékařská fakulta
ISBN
80-210-3053-4
Klíčová slova anglicky
transcriptome; Treponema pallidum; gene expression profiling
Změněno: 31. 5. 2005 09:36, prof. MUDr. David Šmajs, Ph.D.
V originále
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.
Anglicky
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.
Návaznosti
| NI7351, projekt VaV |
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