CBM 4082 PDF

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We have a team of certified expert technicians that can handle any job. Received 26th AprilAccepted 11th July Commonly used phage reporters include alkaline phosphatase phoA0482 beta-galactosidase lacZ28,30,31 green fluorescent protein gfp32,33 and bacterial luciferases luxAB or luxCDABE34—36 among others.

D Schematic of the novel reporter fusion displaying binding affinity to cellulose. Our results suggest that our novel recombinant phages are ideal for an ultrasensitive bacterial detection assay. Reporter bacteriophages phages are robust biorecognition elements uniquely suited for the rapid and sensitive detection of bacterial species. The authors would like to acknowledge additional cmb from the U. In this study, the T7 coliphage was genetically engineered to express the newly developed luceriferase, NanoLuc NLucas an indicator of bacterial cgm.

Upon infection with chm phages, target bacteria express reporter enzymes encoded within the phage genome. The NanoLuc reporter enzyme expression cassette was synthesized as a linear double stranded DNA molecule with 50 bp of phage homology at each terminus. Back to tab navigation Download options Please wait Rapid detection of bacteria responsible for foodborne diseases is a growing necessity for public health. Please enable JavaScript to access the full features of the site or access our non-JavaScript page.


For reproduction of material from NJC: The use of cellulose to collect the reporter probe from the bulk sample solution served to reduce the overall sample volume by over 2 orders of magnitude. Within the range tested, increasing E. For reproduction of material from all other RSC journals: The first time, every time. The advantages of phages include their host specificity, ability to distinguish viable and non-viable cells, low cost, and ease of genetic engineering.

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Positive plaques were isolated, propagated and submitted for full genome sequencing. We have shown the possibility of detecting the immobilized fusion protein in a filter plate which resulted from a single CFU of E.

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Data points represent the average of three replicates and error bars represent the standard deviation. Information about reproducing material from RSC articles with different licences is available on our 40822 Requests page. A current limitation to phage-based assays remains the need for cocktails of phages in order to cover the desired host range. You do not have JavaScript enabled. After an incubation period, the phage and cellulose are added to the sample and the infection incubation period begins, during which expression of NanoLuc-CBM occurs.


This is due to the high specificity of the phages which can often be limited to single strains.

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Talbert c and S. The optical density briefly increased following the addition of reporter phage indicating steady state growth then rapidly dropped as cells lysed. While a cocktail of phages may be required for the detection of an entire bacterial species, we targeted E.

Ongoing work by the authors and other labs aims to customize the tail fibers of phages to allow tailored host ranges. Therefore, we conclude that our phage-based detection assay displays significant aptitude as a proof-of-concept drinking water diagnostic assay for the low-cost, rapid and sensitive detection of E. Hinkley aS. While several genetic approaches were employed to optimize reporter enzyme expression, the novel achievement of this work was the successful fusion of the NanoLuc reporter to a carbohydrate binding module CBM with specificity to crystalline cellulose.