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Related Experiment Videos

Stability of pBR322-derived plasmids.

C S Chiang1, H Bremer

  • 1Biology Programs, University of Texas at Dallas, Richardson 75080.

Plasmid
|November 1, 1988
PubMed
Summary

Plasmid stability in Escherichia coli was assessed without antibiotics. Certain plasmid derivatives showed instability due to cell death and growth advantages of plasmid-free cells, not random partitioning.

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Area of Science:

  • Molecular Biology
  • Microbiology
  • Genetics

Background:

  • Plasmid stability is crucial for maintaining genetic traits in bacterial hosts.
  • The pBR322 plasmid and its derivatives are commonly used vectors in molecular biology.
  • Understanding plasmid loss mechanisms is essential for biotechnological applications.

Purpose of the Study:

  • To investigate the stability of pBR322-derived plasmids in Escherichia coli during growth without antibiotic selection.
  • To differentiate between intrinsic and apparent plasmid instability.
  • To identify factors contributing to plasmid loss.

Main Methods:

  • Culturing Escherichia coli strains harboring various pBR322 derivatives in the absence of antibiotics.
  • Monitoring plasmid retention and loss over multiple bacterial generations.
  • Analyzing plasmid copy number distribution and cell death rates.

Main Results:

  • pBR322, delta rom, and delta bla plasmids were lost within 60 generations.
  • delta tet derivatives exhibited greater stability under the same conditions.
  • Plasmid loss was attributed to a combination of random partitioning and a growth advantage of plasmid-free cells during stationary phase.
  • Cell death, accelerated by plasmid presence and a functional tet gene, contributed to apparent instability.
  • Accumulation of plasmid multimers did not appear to be a significant factor in plasmid instability.

Conclusions:

  • Plasmid instability in E. coli without antibiotics is influenced by both intrinsic random partitioning and apparent factors like cell death and plasmid-free cell growth advantage.
  • The presence of a functional tetracycline resistance gene (tet) on plasmids can decrease their stability.
  • The observed plasmid loss rates deviate significantly from predictions based on random copy number distribution.

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