Construction of mini-Tn10luxABcam/Ptac-ATS and its use for developing a bacteriophage that transduces bioluminescence to Escherichia coli O157:H7

Waddell, Thomas E.

doi:10.1016/s0378-1097(99)00603-5

Cited by 15 publications

(21 citation statements)

References 11 publications

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“…Although phage V10 has been shown to be specific to E. coli O157, it does not plaque on all O157 strains (30). On the other hand, AVR2-V10 killed all 49 O157 isolates tested and may therefore provide complete coverage within the serotype.…”

Section: Discussionmentioning

confidence: 94%

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An Engineered R-Type Pyocin Is a Highly Specific and Sensitive Bactericidal Agent for the Food-Borne Pathogen Escherichia coli O157:H7

Scholl

Cooley

Williams

et al. 2009

Antimicrob Agents Chemother

102

107

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Some strains of Pseudomonas aeruginosa produce R-type pyocins, which are high-molecular-weight phage tail-like protein complexes that have bactericidal activity against other Pseudomonas strains. These particles recognize and bind to bacterial surface structures via tail fibers, their primary spectrum determinant. R-type pyocins kill the cell by contracting a sheath-like structure and inserting their hollow core through the cell envelope, resulting in dissipation of the cellular membrane potential. We have retargeted an R-type pyocin to Escherichia coli O157:H7 by fusing a tail spike protein from an O157-specific phage, V10, to the pyocin tail fiber. The V10 tail spike protein recognizes and degrades the O157 lipopolysaccharide. This engineered pyocin, termed AVR2-V10, is sensitive and specific, killing 100% of diverse E. coli O157:H7 isolates but no other serotypes tested. AVR2-V10 can kill E. coli O157:H7 on beef surfaces, making it a candidate agent for the elimination of this pathogen from food products. All rare AVR2-V10-resistant mutants isolated and examined have lost the ability to produce the O157 antigen and are expected to have compromised virulence. In addition, E. coli O157:H7 exposed to and killed by AVR2-V10 do not release Shiga toxin, as is often the case with many antibiotics, suggesting potential therapeutic applications. The demonstration that a novel R-type pyocin can be created in the laboratory by fusing a catalytic tail spike from the family Podoviridae to a tail fiber of a member of the family Myoviridae is evidence that the plasticity observed among bacteriophage tail genes can, with modern molecular techniques, be exploited to produce nonnatural, targeted antimicrobial agents.

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Section: Discussionmentioning

confidence: 94%

“…Bacteriophage V10 is a phage of the family Podoviridae that specifically infects E. coli O157 strains (30). V10 is similar to P22-like phages, particularly phage ε15 (8,12).…”

Section: Resultsmentioning

confidence: 99%

An Engineered R-Type Pyocin Is a Highly Specific and Sensitive Bactericidal Agent for the Food-Borne Pathogen Escherichia coli O157:H7

Scholl

Cooley

Williams

et al. 2009

Antimicrob Agents Chemother

102

107

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“…In contrast, use of stable chromosomally lux-marked constructs, such as that reported in this study, eliminates the requirement of applying a selective pressure and is thus desirable for long-term survival experiments. Although the construction of a chromosomally lux-marked E. coli O157:H7 strain has been reported previously (41), this was a luxAB mutant, which is dependent on addition of n-decanol for display of the luminescent phenotype. The construct described in this study alleviates this need for additions prior to analysis.…”

Section: Discussionmentioning

confidence: 99%

A Stable Bioluminescent Construct of Escherichia coli O157:H7 for Hazard Assessments of Long-Term Survival in the Environment

Ritchie

Campbell

Shepherd

et al. 2003

Appl Environ Microbiol

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A chromosomally lux-marked (Tn5 luxCDABE) strain of nontoxigenic Escherichia coli O157:H7 was constructed by transposon mutagenesis and shown to have retained the O157, H7, and intimin phenotypes. The survival characteristics of this strain in the experiments performed (soil at ؊5, ؊100, and ؊1,500 kPa matric potential and artificial groundwater) were indistinguishable from the wild-type strain. Evaluation of potential luminescence was found to be a rapid, cheap, and quantitative measure of viable E. coli O157:H7 Tn5 luxCDABE populations in environmental samples. In the survival studies, bioluminescence of the starved populations of E. coli O157:H7 Tn5 luxCDABE could be reactivated to the original levels of light emission, suggesting that these populations remain viable and potentially infective to humans. The attributes of the construct offer a cheap and low-risk substitute to the use of verocytotoxin-producing E. coli O157:H7 in long-term survival studies.

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“…Recently, a renewed interest in phage therapy has surfaced with the advent of antibiotic resistance (3). Phages have also been found useful as diagnostic tools for rapid detection of such pathogens as Mycobacterium tuberculosis, Escherichia coli O157:H7, Listeria monocytogenes, Salmonella spp., and Staphylococcus aureus (5,28,29,36). These diagnostic methods make use of phages that have been genetically modified to carry a reporter gene.…”

mentioning

confidence: 99%

Morphological, Host Range, and Genetic Characterization of Two Coliphages

Goodridge

Gallaccio

Griffiths

2003

Appl Environ Microbiol

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Two coliphages, AR1 and LG1, were characterized based on their morphological, host range, and genetic properties. Transmission electron microscopy showed that both phages belonged to the Myoviridae; phage particles of LG1 were smaller than those of AR1 and had an isometric head 68 nm in diameter and a complex contractile tail 111 nm in length. Transmission electron micrographs of AR1 showed phage particles consisting of an elongated isometric head of 103 by 74 nm and a complex contractile tail 116 nm in length. Both phages were extensively tested on many strains of Escherichia coli and other enterobacteria. The results showed that both phages could infect many serotypes of E. coli. Among the enterobacteria, Proteus mirabilis, Shigella dysenteriae, and two Salmonella strains were lysed by the phages. The genetic material of AR1 and LG1 was characterized. Phage LG1 had a genome size of 49.5 kb compared to 150 kb for AR1. Restriction endonuclease analysis showed that several restriction enzymes could degrade DNA from both phages. The morphological, genome size, and restriction endonuclease similarities between AR1 and phage T4 were striking. Southern hybridizations showed that AR1 and T4 are genetically related. The wide host ranges of phages AR1 and LG1 suggest that they may be useful as biocontrol, therapeutic, or diagnostic agents to control and detect the prevalence of E. coli in animals and food.

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Construction of mini-Tn10luxABcam/Ptac-ATS and its use for developing a bacteriophage that transduces bioluminescence to Escherichia coli O157:H7

Cited by 15 publications

References 11 publications

An Engineered R-Type Pyocin Is a Highly Specific and Sensitive Bactericidal Agent for the Food-Borne Pathogen Escherichia coli O157:H7

An Engineered R-Type Pyocin Is a Highly Specific and Sensitive Bactericidal Agent for the Food-Borne Pathogen Escherichia coli O157:H7

A Stable Bioluminescent Construct of Escherichia coli O157:H7 for Hazard Assessments of Long-Term Survival in the Environment

Morphological, Host Range, and Genetic Characterization of Two Coliphages

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