J 2003

Transcriptome of Treponema pallidum: gene expression profile during experimental rabbit infection

ŠMAJS, David, Petra MATĚJKOVÁ, SJ NORRIS and GM WEINSTOCK

Basic information

Original name

Transcriptome of Treponema pallidum: gene expression profile during experimental rabbit infection

Name in Czech

Transcriptom Treponema pallidum: expresní profilování během experimentální infekce králíků

Authors

ŠMAJS, David (203 Czech Republic, guarantor), Petra MATĚJKOVÁ (203 Czech Republic), SJ NORRIS (840 United States of America) and GM WEINSTOCK (840 United States of America)

Edition

CHEMICKÉ LISTY, Praha, Česká společnost chemická, 2003, 0009-2770

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

Genetics and molecular biology

Country of publisher

Czech Republic

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 0.345

RIV identification code

RIV/00216224:14110/03:00012310

Organization unit

Faculty of Medicine

UT WoS

000227191600037

Keywords in English

transcriptome; Treponema pallidum; gene expression profiling
Změněno: 31/5/2005 10:03, prof. MUDr. David Šmajs, Ph.D.

Abstract

V originále

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.

In Czech

Anotace česky: Byl vyšetřen transkriptom Treponema pallidum během experimentální infekce králíků pomocí DNA microarray čipů obsahujících všech 1039 anotovaných ORF. Geny Treponema pallidum byly rozděleny do skupin podle úrovně transkripce.

Links

NI7351, research and development project
Name: Komparativní genomika patogenních spirochet rodu Treponema: cesta k sekvenčně specifické diagnostice
Investor: Ministry of Health of the CR