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Relationship between plasmid loss and gene expression in Bacillus thuringiensis.

Fatma Driss1, Slim Tounsi, Samir Jaoua

  • 1Laboratory of Biopesticides, Centre of Biotechnology of Sfax, University of Sfax, PO Box 1177, 3018 Sfax, Tunisia.

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Summary

Shortening the Bacillus thuringiensis genome by plasmid loss enhances expression of key genes. This genomic reduction improves chitinolytic, proteolytic, and delta-endotoxin production in mutant strains.

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Area of Science:

  • Microbiology
  • Genomics
  • Molecular Biology

Background:

  • Bacillus thuringiensis possesses a large genome, including essential genes on plasmids.
  • Understanding gene expression regulation in B. thuringiensis is crucial for its applications.

Purpose of the Study:

  • To investigate the impact of genomic reduction via plasmid loss on gene expression in B. thuringiensis.
  • To identify if genome shortening can enhance the production of specific B. thuringiensis proteins.

Main Methods:

  • Provoking plasmid loss in Bacillus thuringiensis to shorten its genome.
  • Analyzing the expression levels of chromosomal and plasmid-borne genes in wild-type and mutant strains.
  • Quantifying chitinolytic, proteolytic, and delta-endotoxin activities in different B. thuringiensis strains.

Main Results:

  • Genomic shortening through plasmid curing led to improved expression of certain chromosomal genes.
  • Totally cured strains (HD1Cry-B, BNS3Cry) exhibited higher chitinolytic and/or proteolytic activities.
  • Partially cured strains (Cur255, CurHD1) showed enhanced delta-endotoxin production (doubled) and bacteriocin activity (1.5-fold increase).

Conclusions:

  • Plasmid loss is an effective strategy for generating B. thuringiensis mutants with enhanced expression of chromosomal genes.
  • Controlled genomic reduction can optimize the production of valuable proteins like delta-endotoxins and bacteriocins.
  • Genome size manipulation offers a viable approach to improve B. thuringiensis strain performance.