Kasurinen , J , Spruit , C M , Wicklund , A , Pajunen , M I & Skurnik , M 2021 , ' Screening of Bacteriophage Encoded Toxic Proteins with a Next Generation Sequencing-Based Assay ' , Viruses (Basel) , vol. 13 , no. 5 , 750 . https://doi.org/10.3390/v13050750
Title: | Screening of Bacteriophage Encoded Toxic Proteins with a Next Generation Sequencing-Based Assay |
Author: | Kasurinen, Jutta; Spruit, Cindy M.; Wicklund, Anu; Pajunen, Maria I.; Skurnik, Mikael |
Contributor organization: | HUMI - Human Microbiome Research Department of Bacteriology and Immunology Medicum Faculty of Medicine University of Helsinki Research Programs Unit HUSLAB Glycoscience Group Biosciences Helsinki One Health (HOH) Mikael Skurnik / Principal Investigator Helsinki University Hospital Area |
Date: | 2021-05 |
Language: | eng |
Number of pages: | 16 |
Belongs to series: | Viruses (Basel) |
ISSN: | 1999-4915 |
DOI: | https://doi.org/10.3390/v13050750 |
URI: | http://hdl.handle.net/10138/332770 |
Abstract: | Bacteriophage vB_EcoM_fHy-Eco03 (fHy-Eco03 for short) was isolated from a sewage sample based on its ability to infect an Escherichia coli clinical blood culture isolate. Altogether, 32 genes encoding hypothetical proteins of unknown function (HPUFs) were identified from the genomic sequence of fHy-Eco03. The HPUFs were screened for toxic properties (toxHPUFs) with a novel, Next Generation Sequencing (NGS)-based approach. This approach identifies toxHPUF-encoding genes through comparison of gene-specific read coverages in DNA from pooled ligation mixtures before electroporation and pooled transformants after electroporation. The performance and reliability of the NGS screening assay was compared with a plating efficiency-based method, and both methods identified the fHy-Eco03 gene g05 product as toxic. While the outcomes of the two screenings were highly similar, the NGS screening assay outperformed the plating efficiency assay in both reliability and efficiency. The NGS screening assay can be used as a high throughput method in the search for new phage-inspired antimicrobial molecules. |
Subject: |
bacteriophage
hypothetical proteins of unknown function next-generation sequencing toxic protein screen ESCHERICHIA-COLI PHAGE SEARCH RECG IDENTIFICATION COMPLEXES DISCOVERY HELICASE GENOMES 3111 Biomedicine 11832 Microbiology and virology |
Peer reviewed: | Yes |
Rights: | cc_by |
Usage restriction: | openAccess |
Self-archived version: | publishedVersion |
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