Determination of copy number of rRNA genes in Pneumocystis carinii f. sp. hominis.

Abstract:

:Differential PCR was performed to determine the copy number of rRNA genes in Pneumocystis carinii f. sp. hominis. Two different reference genes, thymidylate synthase (TS) and beta-tubulin (BTU) genes, were used. Primers for the internal transcribed spacer (ITS) region of nuclear rRNA genes and either the TS or BTU gene were mixed together to perform PCR on seven different bronchoalveolar lavage specimens from patients with P. carinii pneumonia. The radioactivity derived from the incorporated radioactive nucleotides of each PCR product band was then used to calculate the copy number of the ITS relative to that of the TS or BTU gene. The copy number ratio between the ITS and the TS gene was determined to be 0.8, and that between the ITS and the BTU gene was also 0.8. These results suggest that the ITS has the same copy number as the TS or BTU gene. Since the copy number of the TS or BTU gene is presumed to be 1, the results also suggest that P. carinii f. sp. hominis has only one copy of the ITS and thus one copy of the nuclear rRNA genes. Therefore, two types of ITS sequences derived from a specimen would indicate that the patient is infected by two types of P. carinii f. sp. hominis.

journal_name

J Clin Microbiol

authors

Tang X,Bartlett MS,Smith JW,Lu JJ,Lee CH

doi

10.1128/JCM.36.9.2491-2494.1998

subject

Has Abstract

pub_date

1998-09-01 00:00:00

pages

2491-4

issue

9

eissn

0095-1137

issn

1098-660X

journal_volume

36

pub_type

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