A modified in situ RT-PCR method for localizing fungal-specific gene expression in Candida-infected mice renal cells
In situ Reverse Transcriptase PCR (in situ RT-PCR) can amplify mRNA and localize gene expression in cells. However, this method is not feasible in fungi as the thick fungal cell wall constitutes a barrier to this procedure. We developed a two step in situ RT-PCR procedure which enabled the detection...
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| Format: | Article |
| Language: | English |
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Elsevier BV
2009
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| Online Access: | http://psasir.upm.edu.my/id/eprint/12728/1/A%20modified%20in%20situ%20RT-PCR%20method%20for%20localizing%20fungal-specific%20gene%20expression%20in%20Candida-infected%20mice%20renal%20cells.pdf |
| _version_ | 1796968286860083200 |
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| author | Yong, Phelim Voon Chen Ong, Khang Wei Mohd Sidik, Shiran Rosli, Rozita Chong, Pei Pei |
| author_facet | Yong, Phelim Voon Chen Ong, Khang Wei Mohd Sidik, Shiran Rosli, Rozita Chong, Pei Pei |
| author_sort | Yong, Phelim Voon Chen |
| collection | UPM |
| description | In situ Reverse Transcriptase PCR (in situ RT-PCR) can amplify mRNA and localize gene expression in cells. However, this method is not feasible in fungi as the thick fungal cell wall constitutes a barrier to this procedure. We developed a two step in situ RT-PCR procedure which enabled the detection and localization of Candida tropicalis mRNA expression in formalin-fixed, paraffin-embedded (FFPE) mouse kidney sections. This in situ hybridization study revealed the first direct evidence for deposition of Candida tropicalis secreted aspartic proteinase 2 (CtSAP2) in the tip of pseudohyphae and its involvement in acute systemic candidiasis. We conclude that in situ RT-PCR can be successfully applied to FFPE tissues and will offer new perspectives in studying gene expression in Candida species. |
| first_indexed | 2024-03-06T07:26:14Z |
| format | Article |
| id | upm.eprints-12728 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T07:26:14Z |
| publishDate | 2009 |
| publisher | Elsevier BV |
| record_format | dspace |
| spelling | upm.eprints-127282015-11-03T01:45:30Z http://psasir.upm.edu.my/id/eprint/12728/ A modified in situ RT-PCR method for localizing fungal-specific gene expression in Candida-infected mice renal cells Yong, Phelim Voon Chen Ong, Khang Wei Mohd Sidik, Shiran Rosli, Rozita Chong, Pei Pei In situ Reverse Transcriptase PCR (in situ RT-PCR) can amplify mRNA and localize gene expression in cells. However, this method is not feasible in fungi as the thick fungal cell wall constitutes a barrier to this procedure. We developed a two step in situ RT-PCR procedure which enabled the detection and localization of Candida tropicalis mRNA expression in formalin-fixed, paraffin-embedded (FFPE) mouse kidney sections. This in situ hybridization study revealed the first direct evidence for deposition of Candida tropicalis secreted aspartic proteinase 2 (CtSAP2) in the tip of pseudohyphae and its involvement in acute systemic candidiasis. We conclude that in situ RT-PCR can be successfully applied to FFPE tissues and will offer new perspectives in studying gene expression in Candida species. Elsevier BV 2009-11 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/12728/1/A%20modified%20in%20situ%20RT-PCR%20method%20for%20localizing%20fungal-specific%20gene%20expression%20in%20Candida-infected%20mice%20renal%20cells.pdf Yong, Phelim Voon Chen and Ong, Khang Wei and Mohd Sidik, Shiran and Rosli, Rozita and Chong, Pei Pei (2009) A modified in situ RT-PCR method for localizing fungal-specific gene expression in Candida-infected mice renal cells. Journal of Microbiological Methods, 79 (2). pp. 242-245. ISSN 0167-7012; ESSN: 1872-8359 10.1016/j.mimet.2009.08.019 |
| spellingShingle | Yong, Phelim Voon Chen Ong, Khang Wei Mohd Sidik, Shiran Rosli, Rozita Chong, Pei Pei A modified in situ RT-PCR method for localizing fungal-specific gene expression in Candida-infected mice renal cells |
| title | A modified in situ RT-PCR method for localizing fungal-specific gene expression in Candida-infected mice renal cells |
| title_full | A modified in situ RT-PCR method for localizing fungal-specific gene expression in Candida-infected mice renal cells |
| title_fullStr | A modified in situ RT-PCR method for localizing fungal-specific gene expression in Candida-infected mice renal cells |
| title_full_unstemmed | A modified in situ RT-PCR method for localizing fungal-specific gene expression in Candida-infected mice renal cells |
| title_short | A modified in situ RT-PCR method for localizing fungal-specific gene expression in Candida-infected mice renal cells |
| title_sort | modified in situ rt pcr method for localizing fungal specific gene expression in candida infected mice renal cells |
| url | http://psasir.upm.edu.my/id/eprint/12728/1/A%20modified%20in%20situ%20RT-PCR%20method%20for%20localizing%20fungal-specific%20gene%20expression%20in%20Candida-infected%20mice%20renal%20cells.pdf |
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