Cloning and analysis of genomic differences unique to Burkholderia pseudomallei by comparison with B. thailandensis

Brown, Nathaniel Francis; Beacham, Ifor R.

doi:10.1099/0022-1317-49-11-993

Cited by 32 publications

(18 citation statements)

References 31 publications

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“…Manzeniuk et al found that 92% of B. pseudomallei strains produced temperate bacteriophages, demonstrating that prophages are relatively common in this bacterial species (45). Brown and Beacham performed subtractive hybridization between B. pseudomallei and B. thailandensis and identified multiple B. pseudomallei-specific mobile genetic elements, including a P2-like prophage (9). Taken together, the results demonstrate that there is considerable diversity in the mobile genetic elements that B. pseudomallei strains harbor.…”

Section: Discussionmentioning

confidence: 64%

Genomic Diversity of Burkholderia pseudomallei Clinical Isolates: Subtractive Hybridization Reveals a Burkholderia mallei -Specific Prophage in B. pseudomallei 1026b

DeShazer

2004

J Bacteriol

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Burkholderia pseudomallei is the etiologic agent of the disease melioidosis and is a category B biological threat agent. The genomic sequence of B. pseudomallei K96243 was recently determined, but little is known about the overall genetic diversity of this species. Suppression subtractive hybridization was employed to assess the genetic variability between two distinct clinical isolates of B. pseudomallei, 1026b and K96243. Numerous mobile genetic elements, including a temperate bacteriophage designated 1026b, were identified among the 1026b-specific suppression subtractive hybridization products. Bacteriophage 1026b was spontaneously produced by 1026b, and it had a restricted host range, infecting only Burkholderia mallei. It possessed a noncontractile tail, an isometric head, and a linear 54,865-bp genome. The mosaic nature of the 1026b genome was revealed by comparison with bacteriophage E125, a B. mallei-specific bacteriophage produced by Burkholderia thailandensis. The 1026b genes for DNA packaging, tail morphogenesis, host lysis, integration, and DNA replication were nearly identical to the corresponding genes in E125. On the other hand, 1026b genes involved in head morphogenesis were similar to head morphogenesis genes encoded by Pseudomonas putida and Pseudomonas aeruginosa bacteriophages. Consistent with this observation, immunogold electron microscopy demonstrated that polyclonal antiserum against E125 reacted with the tail of 1026b but not with the head. The results presented here suggest that B. pseudomallei strains are genetically heterogeneous and that bacteriophages are major contributors to the genomic diversity of this species. The bacteriophage characterized in this study may be a useful diagnostic tool for differentiating B. pseudomallei and B. mallei, two closely related biological threat agents.

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Section: Discussionmentioning

confidence: 64%

Genomic Diversity of Burkholderia pseudomallei Clinical Isolates: Subtractive Hybridization Reveals a Burkholderia mallei -Specific Prophage in B. pseudomallei 1026b

DeShazer

2004

J Bacteriol

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“…Furthermore, subtractive hybridisation analysis between B. pseudomallei strain 08 and Burkholderia thailandensis, a nonpathogenic relative of B. pseudomallei, indicates that GIs are key determinants of genome plasticity (28). Of 92 sequence fragments found to be present in B. pseudomallei strain 08 and absent from B. thailandensis, only 53 (58%) can be mapped to the K96243 genome.…”

Section: Resultsmentioning

confidence: 99%

Genomic plasticity of the causative agent of melioidosis, Burkholderia pseudomallei

Holden

Titball

Peacock

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

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681

690

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Burkholderia pseudomallei is a recognized biothreat agent and the causative agent of melioidosis. This Gram-negative bacterium exists as a soil saprophyte in melioidosis-endemic areas of the world and accounts for 20% of community-acquired septicaemias in northeastern Thailand where half of those affected die. Here we report the complete genome of B. pseudomallei, which is composed of two chromosomes of 4.07 megabase pairs and 3.17 megabase pairs, showing significant functional partitioning of genes between them. The large chromosome encodes many of the core functions associated with central metabolism and cell growth, whereas the small chromosome carries more accessory functions associated with adaptation and survival in different niches. Genomic comparisons with closely and more distantly related bacteria revealed a greater level of gene order conservation and a greater number of orthologous genes on the large chromosome, suggesting that the two replicons have distinct evolutionary origins. A striking feature of the genome was the presence of 16 genomic islands (GIs) that together made up 6.1% of the genome. Further analysis revealed these islands to be variably present in a collection of invasive and soil isolates but entirely absent from the clonally related organism B. mallei. We propose that variable horizontal gene acquisition by B. pseudomallei is an important feature of recent genetic evolution and that this has resulted in a genetically diverse pathogenic species.

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“…Although important genomic differences exist between the two closely related species B. pseudomallei and B. thailandensis, the genomes of these two species are regarded to be broadly similar (Brown and Beacham, 2000;Yu et al, 2006). Because of this extensive genomic similarity between B. pseudomallei and B. thailandensis, it has been suggested that B. thailandensis could be used as a possible model system for studying certain aspects of B. pseudomallei behaviour (Yu et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

Inflammation patterns induced by different Burkholderia species in mice

Wiersinga¹,

Vos²,

Beer³

et al. 2007

Cell Microbiol

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SummaryBurkholderia pseudomallei, which causes melioidosis, a severe, mainly pulmonary disease endemic in South-East Asia, is considered to be the most pathogenic of the Burkholderia genus. B. thailandensis, however, is considered avirulent. We determined differences in patterns of inflammation of B. pseudomallei 1026b (clinical virulent isolate), B. pseudomallei AJ1D8 (an in vitro invasion-deficient mutant generated from strain 1026b by Tn5-OT182 mutagenesis) and B. thailandensis by intranasally inoculating C57BL/6 mice with each strain. Mice infected with B. thailandensis showed a markedly decreased bacterial outgrowth from lungs, spleen and blood 24 h after inoculation, compared with infection with B. pseudomallei and the invasion mutant AJ1D8. Forty-eight hours after inoculation, B. thailandensis was no longer detectable. This was consistent with elevated pulmonary cytokine and chemokine concentrations after infection with B. pseudomallei 1026b and AJ1D8, and the absence of these mediators 48 h, but not 24 h, after inoculation with B. thailandensis. Histological examination, however, did show marked pulmonary inflammation in the mice infected with B. thailandensis, corresponding with substantial granulocyte influx and raised myeloperoxidase levels. Survival experiments showed that infection with 1 ¥ 10 3 cfu B. thailandensis was not lethal, whereas inoculation with 1 ¥ 10 6 cfu B. thailandensis was equally lethal as 1 ¥ 10 3 cfu B. pseudomallei 1026b or AJ1D8. These data show that B. pseudomallei AJ1D8 is just as lethal as wild-type B. pseudomallei in an in vivo mouse model, and B. thailandensis is perhaps more virulent than is often recognized.

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Cloning and analysis of genomic differences unique to Burkholderia pseudomallei by comparison with B. thailandensis

Cited by 32 publications

References 31 publications

Genomic Diversity of Burkholderia pseudomallei Clinical Isolates: Subtractive Hybridization Reveals a Burkholderia mallei -Specific Prophage in B. pseudomallei 1026b

Genomic Diversity of Burkholderia pseudomallei Clinical Isolates: Subtractive Hybridization Reveals a Burkholderia mallei -Specific Prophage in B. pseudomallei 1026b

Genomic plasticity of the causative agent of melioidosis, Burkholderia pseudomallei

Inflammation patterns induced by different Burkholderia species in mice

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