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Related Concept Videos

Plasmids01:28

Plasmids

Plasmids are extrachromosomal DNA molecules found in bacteria, archaea, and some eukaryotic microbes like yeast. These small, circular DNA structures typically contain fewer than 30 genes, although some may exist linearly. Plasmids vary in their number within a cell, known as copy number. Single-copy plasmids are present in one copy per cell and multi-copy plasmids are present in multiple copies, reaching over 100 copies per cell.Plasmids usually replicate independently of the chromosomal DNA...
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart, a...
Genome Copying Errors02:46

Genome Copying Errors

DNA replication is a well-evolved process that copies millions of base pairs with high fidelity during each cell division. Occasionally a wrong base or a long stretch of wrong bases may get added to the daughter strands. If the errors are left unchecked, cells might accumulate several mutations that might endanger their  survival. Therefore, the copying errors are checked and repaired at three levels.
Translesion DNA Polymerases02:10

Translesion DNA Polymerases

Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
The Replisome03:01

The Replisome

DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...

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Quantification of Plasmid-Mediated Antibiotic Resistance in an Experimental Evolution Approach
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Quantification of Plasmid-Mediated Antibiotic Resistance in an Experimental Evolution Approach

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Multicopy plasmid stability: revisiting the dimer catastrophe.

C M Field1, D K Summers

  • 1Department of Genetics, University of Cambridge, Downing Street, Cambridge CB2 3EH, United Kingdom. cmf40@cam.ac.uk

Journal of Theoretical Biology
|September 28, 2011
PubMed
Summary
This summary is machine-generated.

Bacterial plasmid ColE1 stability is threatened by dimers, but resolution and Rcd checkpoints primarily reduce metabolic burden, not ensure stability. This simulation reveals dimers over-replicate, impacting host cells.

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Plasmid Stability Analysis with Open-Source Droplet Microfluidics

Published on: December 27, 2024

Area of Science:

  • Bacterial genetics
  • Molecular biology
  • Systems biology

Background:

  • Multicopy plasmid ColE1 replication and distribution are crucial for bacterial populations.
  • Plasmid dimers, formed via recombination, threaten high copy numbers and are linked to the 'dimer catastrophe'.
  • The cer site and Rcd RNA regulate dimer resolution and cell division.

Purpose of the Study:

  • To develop a stochastic simulation of ColE1 plasmid replication and distribution.
  • To investigate the role of copy number variance and the Rcd checkpoint in plasmid stability.
  • To re-evaluate the impact of dimers on plasmid stability and host metabolic load.

Main Methods:

  • Constructed a stochastic simulation of ColE1 replication and distribution in growing bacterial cells.
  • Incorporated copy number variance into the dimer catastrophe model.
  • Analyzed the necessity of the Rcd checkpoint under varying dimer resolution rates.

Main Results:

  • Dimers over-replicate compared to monomers, indicating a mechanism for increased metabolic load.
  • The Rcd checkpoint is essential when dimer resolution rates are slow.
  • The impact of dimers on plasmid stability is less severe than previously modeled.

Conclusions:

  • The primary role of dimer resolution and the Rcd checkpoint is to mitigate metabolic burden in recombinogenic hosts.
  • Plasmid stability is less dependent on these mechanisms than initially proposed.
  • Stochastic simulations provide insights into bacterial plasmid dynamics and host-plasmid interactions.