A Toxicity Screening Approach to Identify Bacteriophage-Encoded Anti-Microbial Proteins
| Autor: | Ushanandini Mohanraj, Xing Wan, Mikael Skurnik, Cindy M. Spruit, Maria Pajunen |
|---|---|
| Přispěvatelé: | Medicum, Human Parvoviruses: Epidemiology, Molecular Biology and Clinical Impact, Department of Virology, Antimicrobials, probiotics and fermented food, Department of Bacteriology and Immunology, Helsinki One Health (HOH), Mikael Skurnik / Principal Investigator, HUSLAB, Helsinki University Hospital Area, HUMI - Human Microbiome Research, Glycoscience Group, University Management |
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Models Molecular Proteomics bacteriophages Protein Conformation Cloning vector Genomics toxic medicine.disease_cause Article Microbiology Bacteriophage 03 medical and health sciences Structure-Activity Relationship Viral Proteins HPUF Antibiotic resistance Bacteriolysis Virology medicine Escherichia coli Amino Acid Sequence 1183 Plant biology microbiology virology antibacterials 030102 biochemistry & molecular biology biology Bacteria screening φR1-RT Pathogenic bacteria assay biology.organism_classification Virology & Molecular Biology Virologie & Moleculaire Biologie 030104 developmental biology Infectious Diseases |
| Zdroj: | Viruses 11 (2019) 11 Viruses Volume 11 Issue 11 Viruses, 11(11) |
| ISSN: | 1999-4915 |
| Popis: | The rapid emergence of antibiotic resistance among many pathogenic bacteria has created a profound need to discover new alternatives to antibiotics. Bacteriophages, the viruses of microbes, express special proteins to overtake the metabolism of the bacterial host they infect, the best known of which are involved in bacterial lysis. However, the functions of majority of bacteriophage encoded gene products are not known, i.e., they represent the hypothetical proteins of unknown function (HPUFs). In the current study we present a phage genomics-based screening approach to identify phage HPUFs with antibacterial activity with a long-term goal to use them as leads to find unknown targets to develop novel antibacterial compounds. The screening assay is based on the inhibition of bacterial growth when a toxic gene is expression-cloned into a plasmid vector. It utilizes an optimized plating assay producing a significant difference in the number of transformants after ligation of the toxic and non-toxic genes into a cloning vector. The screening assay was first tested and optimized using several known toxic and non-toxic genes. Then, it was applied to screen 94 HPUFs of bacteriophage &phi R1-RT, and identified four HPUFs that were toxic to Escherichia coli. This optimized assay is in principle useful in the search for bactericidal proteins of any phage, and also opens new possibilities to understanding the strategies bacteriophages use to overtake bacterial hosts. |
| Databáze: | OpenAIRE |
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