1 / 8

Dispersing biofilms with engineered enzymatic bacteriophage

Dispersing biofilms with engineered enzymatic bacteriophage. Doug Tischer. The Problem with Biofilms. Increases resistant to antibiotics Pose a problem for cleaning medical devices, water pipe, food contamination from industrial devices etc. EPS (Extracellular Polymeric Substance)

lisamoon
Download Presentation

Dispersing biofilms with engineered enzymatic bacteriophage

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Dispersing biofilms with engineeredenzymatic bacteriophage Doug Tischer

  2. The Problem with Biofilms • Increases resistant to antibiotics • Pose a problem for cleaning medical devices, water pipe, food contamination from industrial devices etc • EPS (Extracellular Polymeric Substance) • Polysaccharides • Proteins • Nucleic Acids • Lipids

  3. The Idea • Adhesin polymeric β-1,6-N-acetyl-D-glucosamine essential for bacterial adhesion to biofilm • DspB degrades polymeric β -1,6-N-acetyl-D-glucosamine • T3 gene 1.2 allows replication in F-plasmid-containing E. coli Figure taken from Lu, 2007

  4. Construction • T7wt • T7DspB • T7control • Φ10 promotor expresses only during infection • 10B is a capsid protein • S-tag easily detected in western blot Figure taken from Lu, 2007

  5. Results Crystal Violet Staining CFU Counts PFU Counts • No replication (no T3 gene 1.2) Inoculation • T3 gene 1.2 greatly enhances phage efficacy in F-plasmid containing E. coli • DspB enhances plaque destruction • Engineered phage is replicating • PFU = Plaque forming unit Figure taken from Lu, 2007

  6. Results cont. Treated Untreated • 4.5 orders of magnitude drop = 99.997% drop in cell count • Scanning Electron Microscope of cell plaques • Significant disruption of plaques Figure taken from Lu, 2007

  7. My thoughts Liked Disliked • Very practical application • Showed phages can be engineered with several genes of interest without hindering replication • Successful proof of principle • Relatively simple construction • No new behavioral tricks • Only protein expression • No use of premade parts

  8. References • Lu TK, Collins JJ (2007) PNAS 104:11197–11202 • Mark McCormick and Robert Mierendorf. “S•Tag: A Multipurpose Fusion Peptide for Recombinant Proteins.” Novagen Inc.

More Related