Analysis of Variation in Tandem Repeats in the Intron of the Major Surface Glycoprotein Expression Site of the Human Form of<i>Pneumocystis carinii</i>

Ma, Liang; Kutty, Geetha; Jia, Qiuyao; Imamichi, Hiromi; Huang, Laurence; Atzori, Chiara; Beckers, Pascal; Groner, Gena; Beard, Charles B.; Kovacs, Joseph A.

doi:10.1086/345721

Cited by 37 publications

(49 citation statements)

References 22 publications

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“…However, we observed different genotypes in BALF and sputum samples taken from one patient during an episode of PCP. In a study by Ma et al (2002), genotype differences were observed in the upstreamconserved sequence (UCS) repeat region as well as in the mt LSU rRNA gene between sputum and BALF samples or between right and left lung BALF obtained from the same patient. This finding supports the hypothesis of heterogeneity and compartmentalization of P. jirovecii genotypes within the lungs (Helweg-Larsen et al, 2001).…”

Section: Discussionmentioning

confidence: 99%

Genotypic variation of Pneumocystis jirovecii isolates in India based on sequence diversity at mitochondrial large subunit rRNA

Gupta

Mirdha

Guleria

et al. 2011

International Journal of Medical Microbiology

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Section: Discussionmentioning

confidence: 99%

Genotypic variation of Pneumocystis jirovecii isolates in India based on sequence diversity at mitochondrial large subunit rRNA

Gupta

Mirdha

Guleria

et al. 2011

International Journal of Medical Microbiology

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“…The detection of the novel tandem repeat pattern (2, 2, 3, 3) was an interesting result. As reported in another study, most P. jirovecii strains contain three or four repeats (Ma et al, 2002;Esteves et al, 2009).…”

Section: Discussionmentioning

confidence: 63%

“…The capillary electrophoresis method offers highresolution of DNA fragments by size, allowing an easy interpretation of the results and detection of a minor population in mixed populations. The presence of multiple repeat patterns in a single clinical sample was also confirmed by denaturing gel electrophoresis analysis (Ma et al, 2002).…”

Section: Discussionmentioning

confidence: 76%

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A comparison of capillary electrophoresis and direct sequencing in upstream conserved sequence region analysis of Pneumocystis jirovecii strains

Jarboui

Mseddi

Sellami

et al. 2013

Journal of Medical Microbiology

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The MSG is encoded by a multi-copy gene family, with an estimated 80 copies per genome, which are clustered in tandem arrays near the telomeres of each chromosome. Pneumocystis can vary its expressed MSG, presumably as a mechanism to avoid the host immune system (Keely & Stringer, 2009;Cornillot et al., 2002;Keely et al., 2005).The MSG gene is located immediately downstream of a region called the upstream conserved sequence (UCS). The UCS is considered to be essential for the expression of MSG and contains a region of 10 nt tandem repeats with variation in the sequence and in the number of repeats (Kutty et al., 2001;Ma et al., 2002; Esteves et al., 2009).The aim of the present study was to analyse the tandem repeats in the UCS intron of MSG by direct sequencing and to develop a simple and rapid capillary electrophoresis method for typing P. jirovecii and for the detection of mixed infection. METHODSThe study was performed on samples from 13 immunocompromised patients, including 10 HIV-positive patients, two kidney transplant patients and one leukaemia patient. These patients were identified as positive after the amplification of the P. jirovecii mtLSUrRNA gene by PCR (Jarboui et al., 2010).Clinical specimens [bronchoalveolar lavage (BAL) and sputum] isolated from these patients were centrifuged at 1500 g for 5 min and the pellet was digested overnight by proteinase K at 56 uC. DNA was extracted using a commercial kit from Qiagen (QIAamp DNA Minikit; Qiagen).PCR amplification. The PCR assay for the mtLSUrRNA gene was carried out in 50 ml, containing 2.5 mM MgCl 2 ; 0.25 mM each of dATP, dTTP, dCTP and dGTP; 0.25 mM of each primer; and 2.5 U of Taq DNA polymerase (Promega).For the amplification of the UCS genomic DNA sequence of the P. jirovecii MSG gene, we used four primers ( Fig. 1

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“…Therefore, it would appear that the predominant CRJE allele has two copies of the 12 bp sequence, and that the 3-copy allele is a natural variant rather than a PCR artifact. Variation of this sort has been seen in the expression sites of other species of Pneumocystis (Sunkin and Stringer, 1996;Ma et al, 2002). Transition mutations were also observed within the P. murina CRJE, but these were very rare, generally occurring in only one sequence out of 84.…”

Section: Mapping the Downstream Border Of The Crje And Assessment Of mentioning

confidence: 88%

Pneumocystis murina MSG gene family and the structure of the locus associated with its transcription

Keely

Linke

Cushion

et al. 2007

Fungal Genetics and Biology

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Analysis of the Pneumocystis murina MSG gene family and expression-site locus showed that, as in Pneumocystis carinii, P. murina MSG genes are arranged in head-to-tail tandem arrays located on multiple chromosomes, and that a variety of MSG genes can reside at the unique P. murina expression site. Located between the P. murina expression site and attached MSG gene is a block of 132 basepairs that is also present at the beginning of MSG genes that are not at the expression site. The center of this sequence block resembles the 28 basepair CRJE of P. carinii, but the block of conserved sequence in P. murina is nearly five times longer than in P. carinii, and much shorter than in P. wakefieldiae. These data indicate that the P. murina expression-site locus has a distinct structure.

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Analysis of Variation in Tandem Repeats in the Intron of the Major Surface Glycoprotein Expression Site of the Human Form ofPneumocystis carinii

Cited by 37 publications

References 22 publications

Genotypic variation of Pneumocystis jirovecii isolates in India based on sequence diversity at mitochondrial large subunit rRNA

Genotypic variation of Pneumocystis jirovecii isolates in India based on sequence diversity at mitochondrial large subunit rRNA

A comparison of capillary electrophoresis and direct sequencing in upstream conserved sequence region analysis of Pneumocystis jirovecii strains

Pneumocystis murina MSG gene family and the structure of the locus associated with its transcription

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