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Plasmids01:28

Plasmids

109
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...
109
DNA Bacteriophages01:26

DNA Bacteriophages

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Bacteriophages, or phages, are viruses that specifically infect bacteria, utilizing their genetic material to hijack host cellular machinery for replication. DNA bacteriophages employ single-stranded DNA (ssDNA) or double-stranded DNA (dsDNA) genomes. These phages exhibit diverse replication strategies and host interactions, influencing their ecological roles and applications in biotechnology and medicine.ssDNA BacteriophagesssDNA phages, with their small genomes, utilize unique strategies to...
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Repressible Operon: trp Operon01:21

Repressible Operon: trp Operon

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The trp operon in Escherichia coli exemplifies a repressible operon. It regulates the synthesis of tryptophan through repressor-mediated transcriptional control and attenuation. This dual regulatory mechanism ensures tryptophan biosynthesis occurs only when needed, conserving cellular resources.Structure of the trp OperonThe trp operon consists of five structural genes (trpE, trpD, trpC, trpB, and trpA) that encode enzymes for tryptophan biosynthesis. These genes are transcribed as a single...
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Related Experiment Video

Updated: Sep 5, 2025

Quantification of Plasmid-Mediated Antibiotic Resistance in an Experimental Evolution Approach
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A plasmid system with tunable copy number.

Miles V Rouches1, Yasu Xu1, Louis Brian Georges Cortes2

  • 1Field of Biophysics, Cornell University, Ithaca, NY, 14853, USA.

Nature Communications
|July 7, 2022
PubMed
Summary
This summary is machine-generated.

Researchers developed new systems for precise control over plasmid copy number in bacteria, enabling fine-tuning of gene expression and synthetic biology optimization. This advancement impacts genetic engineering and recombinant gene expression.

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Area of Science:

  • Synthetic Biology
  • Molecular Biology
  • Bacteriology

Background:

  • Plasmids are essential tools for genetic engineering and recombinant gene expression in bacteria.
  • Current plasmid cloning vectors have limited control over copy number, restricting optimization.
  • Existing Origins of Replication (ORIs) are derived from naturally occurring plasmids.

Purpose of the Study:

  • To introduce novel systems for continuous and fine-tuned control of plasmid copy number.
  • To investigate the impact of varying plasmid copy numbers on cellular processes.
  • To optimize synthetic biological systems through precise gene expression control.

Main Methods:

  • Development of an anhydrotetracycline-controlled plasmid system for tunable copy number.
  • Creation of a parallelized assay to generate a wide spectrum of ColE1-based copy number variants.
  • Utilizing single-cell timelapse measurements to analyze plasmid dynamics and gene expression variability.

Main Results:

  • Achieved continuous control of plasmid copy number from 1 to 800 copies per cell.
  • Demonstrated the effects of plasmid copy number on cellular growth, gene expression, and biosynthesis.
  • Quantified a linear metabolic burden of 0.063% per plasmid, linking it to DNA synthesis.

Conclusions:

  • The developed systems offer precise control over gene expression via plasmid copy number.
  • Tuning plasmid copy number is a critical factor for optimizing synthetic biological systems.
  • The findings suggest a straightforward relationship between metabolic burden and plasmid DNA synthesis.