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

Xing Tang, Marilyn S. Bartlett, James W. Smith, Jang Jih Lu, Chao Hung Lee

Research output: Contribution to journalArticle

36 Citations (Scopus)

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 gent 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.

Original languageEnglish (US)
Pages (from-to)2491-2494
Number of pages4
JournalJournal of clinical microbiology
Volume36
Issue number9
StatePublished - Sep 1 1998

Fingerprint

Pneumocystis carinii
Thymidylate Synthase
rRNA Genes
Tubulin
Genes
Polymerase Chain Reaction
Pneumocystis Pneumonia
Bronchoalveolar Lavage
Radioactivity
Nucleotides

ASJC Scopus subject areas

  • Microbiology (medical)

Cite this

Tang, X., Bartlett, M. S., Smith, J. W., Lu, J. J., & Lee, C. H. (1998). Determination of copy number of rRNA genes in Pneumocystis carinii f. sp. hominis. Journal of clinical microbiology, 36(9), 2491-2494.

Determination of copy number of rRNA genes in Pneumocystis carinii f. sp. hominis. / Tang, Xing; Bartlett, Marilyn S.; Smith, James W.; Lu, Jang Jih; Lee, Chao Hung.

In: Journal of clinical microbiology, Vol. 36, No. 9, 01.09.1998, p. 2491-2494.

Research output: Contribution to journalArticle

Tang, X, Bartlett, MS, Smith, JW, Lu, JJ & Lee, CH 1998, 'Determination of copy number of rRNA genes in Pneumocystis carinii f. sp. hominis', Journal of clinical microbiology, vol. 36, no. 9, pp. 2491-2494.
Tang X, Bartlett MS, Smith JW, Lu JJ, Lee CH. Determination of copy number of rRNA genes in Pneumocystis carinii f. sp. hominis. Journal of clinical microbiology. 1998 Sep 1;36(9):2491-2494.
Tang, Xing ; Bartlett, Marilyn S. ; Smith, James W. ; Lu, Jang Jih ; Lee, Chao Hung. / Determination of copy number of rRNA genes in Pneumocystis carinii f. sp. hominis. In: Journal of clinical microbiology. 1998 ; Vol. 36, No. 9. pp. 2491-2494.
@article{5be3bcee50d748b693b03d906a46a069,
title = "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 gent 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.",
author = "Xing Tang and Bartlett, {Marilyn S.} and Smith, {James W.} and Lu, {Jang Jih} and Lee, {Chao Hung}",
year = "1998",
month = "9",
day = "1",
language = "English (US)",
volume = "36",
pages = "2491--2494",
journal = "Journal of Clinical Microbiology",
issn = "0095-1137",
publisher = "American Society for Microbiology",
number = "9",

}

TY - JOUR

T1 - Determination of copy number of rRNA genes in Pneumocystis carinii f. sp. hominis

AU - Tang, Xing

AU - Bartlett, Marilyn S.

AU - Smith, James W.

AU - Lu, Jang Jih

AU - Lee, Chao Hung

PY - 1998/9/1

Y1 - 1998/9/1

N2 - 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 gent 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.

AB - 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 gent 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.

UR - http://www.scopus.com/inward/record.url?scp=0031850247&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031850247&partnerID=8YFLogxK

M3 - Article

C2 - 9705380

AN - SCOPUS:0031850247

VL - 36

SP - 2491

EP - 2494

JO - Journal of Clinical Microbiology

JF - Journal of Clinical Microbiology

SN - 0095-1137

IS - 9

ER -