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Published on: April 9, 2015
Isolation and characterization of bacteriophage PhiBP from Paenibacillus polymyxa CCM 7400
Nora Halgasova1, Jana Ugorcakova, Martina Gerova
1Institute of Molecular Biology, Slovak Academy of Sciences, Bratislava, Slovakia.
Abstract:
A bacteriophage PhiBP infecting Paenibacillus polymyxa CCM 7400 was isolated from culture lysate. Electron microscopy of lysate samples revealed the presence of bacteriophage particles with polyhedral heads 56 nm in diameter and flexible noncontractile tails 144 nm in length. The profile of PhiBP structural proteins resembles that of other bacteriophages. The PhiBP genome consists of double-stranded DNA of 43-kbp size. Homology search of sequenced DNA fragments from EcoRI digest revealed regions with significant similarity to other known bacteriophage genes. Regions similar to phage terminase genes were identified within the 1.2-kbp fragment. Three lytic genes, two holin genes and one endolysin gene were identified within the 2.5-kbp fragment. We tested the isolates of P. polymyxa CCM 7400 for the presence of phage DNA on bacterial chromosome using PCR amplification with primers derived from proposed terminase and holin gene sequences. We confirmed the presence of PhiBP DNA on P. polymyxa chromosome by Southern hybridization. The bacteriophage PhiBP was capable of causing lysis of a P. polymyxaPhiBP lysogen despite the presence of the phage DNA on bacterial chromosome. Therefore, we concluded that PhiBP was a virulent mutant phage.
Insights
This study isolated and characterized bacteriophage PhiBP, a virulent mutant of Paenibacillus polymyxa. PhiBP DNA was found integrated into the bacterial chromosome, yet the phage still caused lysis, confirming its virulent nature.
Area of Science:
- Microbiology
- Virology
- Molecular Biology
Background:
- Paenibacillus polymyxa is a bacterium with agricultural and industrial applications.
- Bacteriophages are viruses that infect bacteria and play roles in microbial ecology.
- Understanding phage-host interactions is crucial for phage therapy and biotechnology.
Purpose of the Study:
- To isolate and characterize bacteriophage PhiBP infecting Paenibacillus polymyxa CCM 7400.
- To determine the genomic and structural properties of PhiBP.
- To investigate the virulence of PhiBP, particularly its ability to lyse lysogenic strains.
Main Methods:
- Isolation and purification of bacteriophage PhiBP from bacterial culture lysate.
- Electron microscopy for structural analysis of phage particles.
- Genomic DNA extraction, restriction digestion (EcoRI), and DNA sequencing.
- Bioinformatic analysis for homology search and gene identification (terminase, holin, endolysin).
- Polymerase Chain Reaction (PCR) and Southern hybridization to detect phage DNA integration in the bacterial chromosome.
Main Results:
- Bacteriophage PhiBP possesses polyhedral heads (56 nm) and flexible tails (144 nm).
- The PhiBP genome is double-stranded DNA (43-kbp) with homology to known phage genes, including terminase, holin, and endolysin.
- PhiBP DNA was confirmed to be integrated into the Paenibacillus polymyxa CCM 7400 chromosome.
- PhiBP successfully lysed a Paenibacillus polymyxa CCM 7400 lysogen, indicating virulent behavior.
Conclusions:
- PhiBP is a virulent mutant bacteriophage infecting Paenibacillus polymyxa.
- The presence of integrated phage DNA did not confer immunity to lysis, a characteristic of virulent phages.
- This finding has implications for understanding phage-bacterial interactions and potential applications in controlling Paenibacillus polymyxa populations.
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