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Molecular Characterization of Putative Virulence Determinants in Burkholderia pseudomallei
The Gram-negative saprophyte Burkholderia pseudomallei is the causative agent of melioidosis, an infectious disease which is endemic in Southeast Asia and northern Australia. This bacterium possesses many virulence factors which are thought to contribute to its survival and pathogenicity. Using a vi...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Hindawi Publishing Corporation
2014
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4158159/ https://www.ncbi.nlm.nih.gov/pubmed/25215325 http://dx.doi.org/10.1155/2014/590803 |
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author | Puah, Suat Moi Puthucheary, S. D. Wang, Jin Town Pan, Yi Jiun Chua, Kek Heng |
author_facet | Puah, Suat Moi Puthucheary, S. D. Wang, Jin Town Pan, Yi Jiun Chua, Kek Heng |
author_sort | Puah, Suat Moi |
collection | PubMed |
description | The Gram-negative saprophyte Burkholderia pseudomallei is the causative agent of melioidosis, an infectious disease which is endemic in Southeast Asia and northern Australia. This bacterium possesses many virulence factors which are thought to contribute to its survival and pathogenicity. Using a virulent clinical isolate of B. pseudomallei and an attenuated strain of the same B. pseudomallei isolate, 6 genes BPSL2033, BP1026B_I2784, BP1026B_I2780, BURPS1106A_A0094, BURPS1106A_1131, and BURPS1710A_1419 were identified earlier by PCR-based subtractive hybridization. These genes were extensively characterized at the molecular level, together with an additional gene BPSL3147 that had been identified by other investigators. Through a reverse genetic approach, single-gene knockout mutants were successfully constructed by using site-specific insertion mutagenesis and were confirmed by PCR. BPSL2033::Km and BURPS1710A_1419::Km mutants showed reduced rates of survival inside macrophage RAW 264.7 cells and also low levels of virulence in the nematode infection model. BPSL2033::Km demonstrated weak statistical significance (P = 0.049) at 8 hours after infection in macrophage infection study but this was not seen in BURPS1710A_1419::Km. Nevertheless, complemented strains of both genes were able to partially restore the gene defects in both in vitro and in vivo studies, thus suggesting that they individually play a minor role in the virulence of B. pseudomallei. |
format | Online Article Text |
id | pubmed-4158159 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Hindawi Publishing Corporation |
record_format | MEDLINE/PubMed |
spelling | pubmed-41581592014-09-11 Molecular Characterization of Putative Virulence Determinants in Burkholderia pseudomallei Puah, Suat Moi Puthucheary, S. D. Wang, Jin Town Pan, Yi Jiun Chua, Kek Heng ScientificWorldJournal Research Article The Gram-negative saprophyte Burkholderia pseudomallei is the causative agent of melioidosis, an infectious disease which is endemic in Southeast Asia and northern Australia. This bacterium possesses many virulence factors which are thought to contribute to its survival and pathogenicity. Using a virulent clinical isolate of B. pseudomallei and an attenuated strain of the same B. pseudomallei isolate, 6 genes BPSL2033, BP1026B_I2784, BP1026B_I2780, BURPS1106A_A0094, BURPS1106A_1131, and BURPS1710A_1419 were identified earlier by PCR-based subtractive hybridization. These genes were extensively characterized at the molecular level, together with an additional gene BPSL3147 that had been identified by other investigators. Through a reverse genetic approach, single-gene knockout mutants were successfully constructed by using site-specific insertion mutagenesis and were confirmed by PCR. BPSL2033::Km and BURPS1710A_1419::Km mutants showed reduced rates of survival inside macrophage RAW 264.7 cells and also low levels of virulence in the nematode infection model. BPSL2033::Km demonstrated weak statistical significance (P = 0.049) at 8 hours after infection in macrophage infection study but this was not seen in BURPS1710A_1419::Km. Nevertheless, complemented strains of both genes were able to partially restore the gene defects in both in vitro and in vivo studies, thus suggesting that they individually play a minor role in the virulence of B. pseudomallei. Hindawi Publishing Corporation 2014 2014-08-11 /pmc/articles/PMC4158159/ /pubmed/25215325 http://dx.doi.org/10.1155/2014/590803 Text en Copyright © 2014 Suat Moi Puah et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Puah, Suat Moi Puthucheary, S. D. Wang, Jin Town Pan, Yi Jiun Chua, Kek Heng Molecular Characterization of Putative Virulence Determinants in Burkholderia pseudomallei |
title | Molecular Characterization of Putative Virulence Determinants in Burkholderia pseudomallei
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title_full | Molecular Characterization of Putative Virulence Determinants in Burkholderia pseudomallei
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title_fullStr | Molecular Characterization of Putative Virulence Determinants in Burkholderia pseudomallei
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title_full_unstemmed | Molecular Characterization of Putative Virulence Determinants in Burkholderia pseudomallei
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title_short | Molecular Characterization of Putative Virulence Determinants in Burkholderia pseudomallei
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title_sort | molecular characterization of putative virulence determinants in burkholderia pseudomallei |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4158159/ https://www.ncbi.nlm.nih.gov/pubmed/25215325 http://dx.doi.org/10.1155/2014/590803 |
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