ŠMAJS, David, Petra MATĚJKOVÁ, SJ NORRIS and GM WEINSTOCK. Transcriptome of Treponema pallidum: gene expression profiling of treponemes grown in rabbits. In Sborník příspěvků VII. Pracovní setkání biochemiků a molekulárních biologů. 2003. vyd. Brno: Masarykova univerzita, 2003. p. 12. ISBN 80-210-3053-4.
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Basic information
Original name Transcriptome of Treponema pallidum: gene expression profiling of treponemes grown in rabbits
Name in Czech Transcriptom Treponema pallidum: expresní profilování během experimentální infekce králíků
Name (in English) Transcriptome of Treponema pallidum: gene expression profiling of treponemes grown in rabbits
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 2003. vyd. Brno, Sborník příspěvků VII. Pracovní setkání biochemiků a molekulárních biologů, p. 12-12, 2003.
Publisher Masarykova univerzita
Other information
Original language Czech
Type of outcome Proceedings paper
Field of Study Genetics and molecular biology
Country of publisher Czech Republic
Confidentiality degree is not subject to a state or trade secret
RIV identification code RIV/00216224:14110/03:00012307
Organization unit Faculty of Medicine
ISBN 80-210-3053-4
Keywords in English transcriptome; Treponema pallidum; gene expression profiling
Tags gene expression profiling, transcriptome, Treponema pallidum
Changed by Changed by: prof. MUDr. David Šmajs, Ph.D., učo 1116. Changed: 31/5/2005 09:36.
Abstract
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.
Abstract (in English)
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.
Links
NI7351, research and development projectName: Komparativní genomika patogenních spirochet rodu Treponema: cesta k sekvenčně specifické diagnostice
Investor: Ministry of Health of the CR, Infectious Diseases, Microbiology, Epidemiology and Immunology
PrintDisplayed: 31/3/2020 16:50