Determination of the copy number of the nuclear rDNA and beta-tubulin genes of Pneumocystis carinii f. sp. hominis using PCR multicompetitors

Details

Serval ID
serval:BIB_E54F8EBD3D9D
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Determination of the copy number of the nuclear rDNA and beta-tubulin genes of Pneumocystis carinii f. sp. hominis using PCR multicompetitors
Journal
Journal of Eukaryotic Microbiology
Author(s)
Nahimana  A., Francioli  P., Blanc  D. S., Bille  J., Wakefield  A. E., Hauser  P. M.
ISSN
1066-5234
Publication state
Published
Issued date
08/2000
Peer-reviewed
Oui
Volume
47
Number
4
Pages
368-72
Notes
Evaluation Studies
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Jul-Aug
Abstract
Multiple copies of a gene may lead to difficulty in the interpretation of typing results because polymorphism of the copies may wrongly lead to the conclusion that different types are present in a specimen. To determine the copy number per genome of the nuclear rDNA and beta-tubulin genes analyzed for the typing of Pneumocystis carinii f. sp. hominis, we developed a strategy based on the use of the same multicompetitor molecule in two different quantitative-competitive PCRs, one for the gene under study and the other for a reference single copy gene, allowing direct comparison of the results of both PCRs. Control experiments showed that the strategy was sensitive enough to detect duplication of a gene. The copy number of the nuclear rDNA operon was determined by amplification of the intron of the 26S rDNA gene and that of the beta-tubulin by amplification of the region surrounding the intron no. 6. The method was first tested on P. c. carinii, the special form commonly infecting rats. Pneumocystis c. carinii was found to contain a single copy of the rDNA operon. The method was then applied to P. c. hominis. The results confirmed that P. c. hominis genome contains a single copy of the nuclear rDNA and beta-tubulin genes.
Keywords
AIDS-Related Opportunistic Infections/microbiology Animals Bronchoalveolar Lavage Cell Nucleus/genetics DNA, Ribosomal/*genetics *Gene Dosage Humans Pneumocystis/*genetics Pneumonia, Pneumocystis/microbiology Polymerase Chain Reaction/*methods Rats Sensitivity and Specificity Tubulin/*genetics
Pubmed
Web of science
Create date
29/01/2008 15:20
Last modification date
20/08/2019 16:08
Usage data