Escherichia coli detection by GFP-labeled lysozyme-inactivated T4 bacteriophage

Tanji, Yasunori; Furukawa, Chiaki; Na, Suk-Hyun; Hijikata, Tomonori; Miyanaga, Kazuhiko; Unno, Hajime

doi:10.1016/j.jbiotec.2004.05.011

Cited by 72 publications

(59 citation statements)

References 21 publications

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“…20 Similarly, several researchers combined the reporter phage concept and labeling of phages to create genetically modified labeled phages, which displayed the green fluorescent reporter protein on the capsid surface. 21,22 This technique allowed the detection of viable but non-culturable bacteria (VBNC) within one hour, if sufficient levels of bacteria were present. 21,22 Phages have also been labeled with quantum dots and used to detect bacteria.…”

Section: Resultsmentioning

confidence: 99%

“…21,22 This technique allowed the detection of viable but non-culturable bacteria (VBNC) within one hour, if sufficient levels of bacteria were present. 21,22 Phages have also been labeled with quantum dots and used to detect bacteria. 23 One disadvantage of the above studies is the fact that they all required expensive instrumentation to detect the fluorescently phage-based methods have been developed to effect rapid detection of bacteria.…”

Section: Resultsmentioning

confidence: 99%

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Luminescence based enzyme-labeled phage (Phazyme) assays for rapid detection of Shiga toxin producingEscherichia coliserogroups

et al. 2011

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Luminescence based enzyme-labeled phage (Phazyme) assays for rapid detection of Shiga toxin producingEscherichia coliserogroups

et al. 2011

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“…Without an efficient phage screening method, finding the desired clone is labor-intensive at best. Therefore, a reporter gene, usually encoding luciferase or a fluorescent protein, is commonly cloned along with the gene of interest to facilitate the identification of mutant phages by detecting the reporter (59,(62)(63)(64)(65). Because the recombination rates obtained with this technique are low, it is improbable that targeting multiple loci at the same time will result in an organism carrying all the desired mutations.…”

Section: Techniques For Engineering Synthetic Phages Homologous Recommentioning

confidence: 99%

“…Phages expressing green fluorescent protein (GFP) have been proposed as a fast and accurate method for detecting E. coli (64,65,174). The gfp gene, originally carried on a plasmid, was inserted by homologous recombination into the genomes of phages T4 (wild type), T4e…”

Section: Phage Engineering For Bacterial Detection and Diagnosticsmentioning

confidence: 99%

“…Ϫ (a gene e amber mutant phage) (64), and PP01 (65) such that it was displayed on the small outer capsid (SOC) of these phages, resulting in phages T4wt/GFP, T4e Ϫ /GFP, and PP01-SOC/GFP (GFP introduced into the C terminus of SOC) or PP01-GFP/SOC (GFP introduced into the N terminus of SOC), respectively. The gfp gene was also inserted into phages IP008 and IP052, within the e gene, which encodes a phage lysozyme, resulting in phages IP008e-/GFP and IP052e-/GFP, which exhibited suppressed lytic activity (174).…”

Section: Phage Engineering For Bacterial Detection and Diagnosticsmentioning

confidence: 99%

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Genetically Engineered Phages: a Review of Advances over the Last Decade

Pires

Cleto

Sillankorva

et al. 2016

Microbiol Mol Biol Rev

336

275

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SUMMARYSoon after their discovery in the early 20th century, bacteriophages were recognized to have great potential as antimicrobial agents, a potential that has yet to be fully realized. The nascent field of phage therapy was adversely affected by inadequately controlled trials and the discovery of antibiotics. Although the study of phages as anti-infective agents slowed, phages played an important role in the development of molecular biology. In recent years, the increase in multidrug-resistant bacteria has renewed interest in the use of phages as antimicrobial agents. With the wide array of possibilities offered by genetic engineering, these bacterial viruses are being modified to precisely control and detect bacteria and to serve as new sources of antibacterials. In applications that go beyond their antimicrobial activity, phages are also being developed as vehicles for drug delivery and vaccines, as well as for the assembly of new materials. This review highlights advances in techniques used to engineer phages for all of these purposes and discusses existing challenges and opportunities for future work.

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IgA response of BALB/c mice to orally administered Salmonella typhimurium flagellin‐displaying T2 bacteriophages

Synnott

Ohshima

Nakai

et al. 2009

Biotechnology Progress

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Salmonella typhimurium antigens were displayed on the capsid of a T2 bacteriophage to explore the potential of phage display for an oral vaccine. Segments of the flagellin proteins FliC (H1 antigen) and FljB (H2) were fused to the N-terminal of T2 phage SOC to give two recombinant phages, T2FliCm and T2FljBm. Over 14 days, 19 BALB/c mice were orally administered twice, either with purified recombinant FliCm and FljBm protein, or T2FliCm and T2FljBm with or without host Escherichia coli. Feces were sampled over 10 weeks and examined for phage by plaque assay and for the presence of mucosal IgA by ELISA. Relatively few phages were detected relative to the amount administered (up to 8.21 x 10(3) PFU/g faeces) and none were detected five days after initial administration. The administration of a large number of phages appeared to cause no clinical symptoms. IgA concentration in feces peaked around four weeks after the second administration and subsided after eight weeks. The highest relative titers were observed in the protein group (0.37% for anti-FliCm and 0.22% for anti-FljBm) and the mouse group which received no E. coli (0.33% and 0.35%) despite the theoretical amount of protein contained in a phage dose being at least 80-465 times lower than the protein dose administered. The possibility that the immuno-stimulatory properties of the phage create an adjuvant effect to enhance the immunogenic properties of the displayed proteins is discussed. We conclude that phage may be valuable as a vector for oral vaccines.

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Escherichia coli detection by GFP-labeled lysozyme-inactivated T4 bacteriophage

Cited by 72 publications

References 21 publications

Luminescence based enzyme-labeled phage (Phazyme) assays for rapid detection of Shiga toxin producingEscherichia coliserogroups

Luminescence based enzyme-labeled phage (Phazyme) assays for rapid detection of Shiga toxin producingEscherichia coliserogroups

Genetically Engineered Phages: a Review of Advances over the Last Decade

IgA response of BALB/c mice to orally administered Salmonella typhimurium flagellin‐displaying T2 bacteriophages

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