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Genetic Engineering of Lysogenic-Lytic Switch Genes Improves Burkholderia Phage Killing Efficacy

Pacharapong Khrongsee1,2, Sarah M Doore3, Nawarat Somprasong4

  • 1Department of Infectious Diseases and Immunology, College of Veterinary Medicine, University of Florida, Gainesville, FL 32608, USA.

International Journal of Molecular Sciences
|March 28, 2026
PubMed

Insights

Temperate phages from Burkholderia pseudomallei, the cause of melioidosis, can be engineered for improved lytic activity. Deleting specific regulatory genes in phage φBP82.2 enhances its killing capabilities, offering new therapeutic strategies.

Area of Science:

  • Microbiology
  • Bacteriology
  • Virology

Background:

  • Burkholderia pseudomallei causes melioidosis, a disease with difficult treatment and decontamination.
  • Bacteriophages (phages) are promising biocontrol agents against B. pseudomallei.
  • Prophages within B. pseudomallei genomes are potential sources of temperate phages.

Purpose of the Study:

  • To investigate the lysogenic-lytic control mechanisms of temperate phages derived from B. pseudomallei.
  • To identify genes involved in the lysogenic-lytic switch of the P2-like phage φBP82.2.
  • To explore engineering strategies for enhanced phage lytic activity against B. pseudomallei.

Main Methods:

  • Transcriptomic profiling of B. pseudomallei Bp82 to identify highly expressed phage genes.
  • Genetic manipulation of temperate phage φBP82.2, including integrase gene mutagenesis and deletion of regulatory genes.
  • Comparative analysis of wildtype and engineered phage killing activity.

Main Results:

  • High expression of integrase and regulatory genes was observed in prophages during B. pseudomallei growth.
  • Mutagenesis of the integrase gene (phiBP82.2_gp51) did not enhance phage killing activity.
  • Deletion of a putative transcriptional regulatory gene (phiBP82.2_gp38) and downstream genes (phiBP82.2_gp36, phiBP82.2_gp37) significantly improved the lytic activity of phage φBP82.2.

Conclusions:

  • Specific genes (phiBP82.2_gp38, phiBP82.2_gp36, phiBP82.2_gp37) are critical for the lysogenic-lytic switch in temperate phage φBP82.2.
  • Engineering temperate phages by targeting these genes offers a novel approach for developing phage-based therapies against B. pseudomallei.
  • This research opens new avenues for utilizing temperate phages as biocontrol agents for melioidosis treatment and decontamination.
Keywords:
Burkholderia pseudomalleimelioidosisphage engineeringprophage

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