Construction of an inducible system for the analysis of essential genes in Yersinia pestis

Ford, Donna C.; Ireland, Philip M.; Bullifent, Helen L.; Saint, Richard J.; McAlister, Erin V.; Sarkar‐Tyson, Mitali; Oyston, Petra C. F.

doi:10.1016/j.mimet.2014.01.017

Cited by 4 publications

(2 citation statements)

References 42 publications

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“…Targets selected for validation of the essentials list were placed under the control of a rhamnose-inducible promoter system using a method described previously 39 . Briefly, the open reading frame was cloned into the low copy number plasmid vector pBADrha, under the regulatory control of the rhaB gene promoter and accessory genes rhaS/R, before transformation into Y. pestis containing the λ-red recombinase helper plasmid, pAJD434.…”

Section: Methodsmentioning

confidence: 99%

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A Noise Trimming and Positional Significance of Transposon Insertion System to Identify Essential Genes in Yersinia pestis

Yang

Bullifent

Moore

et al. 2017

Sci Rep

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Massively parallel sequencing technology coupled with saturation mutagenesis has provided new and global insights into gene functions and roles. At a simplistic level, the frequency of mutations within genes can indicate the degree of essentiality. However, this approach neglects to take account of the positional significance of mutations - the function of a gene is less likely to be disrupted by a mutation close to the distal ends. Therefore, a systematic bioinformatics approach to improve the reliability of essential gene identification is desirable. We report here a parametric model which introduces a novel mutation feature together with a noise trimming approach to predict the biological significance of Tn5 mutations. We show improved performance of essential gene prediction in the bacterium Yersinia pestis, the causative agent of plague. This method would have broad applicability to other organisms and to the identification of genes which are essential for competitiveness or survival under a broad range of stresses.

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Section: Methodsmentioning

confidence: 99%

“…Bacteria were pelleted to facilitate concentration before addition to BAB broth supplemented with rhamnose or glucose as above to a starting A 590 of 0.06. Growth curves were performed (x6) in a 96-well format using a Multiskan FC incubated plate reader as described previously 39 . Alternatively, essentiality was confirmed on solid media.…”

Section: Methodsmentioning

confidence: 99%

A Noise Trimming and Positional Significance of Transposon Insertion System to Identify Essential Genes in Yersinia pestis

Yang

Bullifent

Moore

et al. 2017

Sci Rep

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Survival protein A is essential for virulence in Yersinia pestis

Southern

Scott

Jenner

et al. 2016

Microbial Pathogenesis

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Plague is a highly pathogenic disease caused by the bacterium Yersinia pestis. There is currently no vaccine available for prophylaxis and antibiotic resistant strains have been isolated, thus there is a need for the development of new countermeasures to treat this disease. Survival protein A (SurA) is a chaperone that has been linked to virulence in several species of bacteria, including the close relative Yersinia pseudotuberculosis. In this study, we aimed to evaluate the role of SurA in virulence of the highly pathogenic Y. pestis by creating an unmarked surA deletion mutant. The Y. pestis ΔsurA mutant was found to be more susceptible to membrane perturbing agents and was completely avirulent in a mouse infection model when delivered up to 2.1 × 10(5) CFU by the subcutaneous route. This provides strong evidence that SurA would make a promising antimicrobial target.

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Proteome and phospholipid alteration reveal metabolic network of Bacillus thuringiensis under triclosan stress

Qin

et al. 2018

Science of The Total Environment

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Construction of an inducible system for the analysis of essential genes in Yersinia pestis

Cited by 4 publications

References 42 publications

A Noise Trimming and Positional Significance of Transposon Insertion System to Identify Essential Genes in Yersinia pestis

A Noise Trimming and Positional Significance of Transposon Insertion System to Identify Essential Genes in Yersinia pestis

Survival protein A is essential for virulence in Yersinia pestis

Proteome and phospholipid alteration reveal metabolic network of Bacillus thuringiensis under triclosan stress

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