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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...

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A biobrick library for cloning custom eukaryotic plasmids.

Marco Constante1, Raik Grünberg, Mark Isalan

  • 1EMBL/CRG Systems Biology Research Unit, Centre for Genomic Regulation (CRG) and UPF, Barcelona, Spain. marco.constante@crg.es

Plos One
|September 9, 2011
PubMed
Summary

Researchers can now build custom plasmids using standard synthetic biology parts (biobricks). This versatile collection enables the creation of novel constructs, including fluorescent reporter plasmids and tools for gene insertion, streamlining genetic engineering workflows.

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Area of Science:

  • Synthetic Biology
  • Molecular Biology
  • Genetic Engineering

Background:

  • Customized plasmids are frequently needed but not commercially available.
  • Standardized biological parts (biobricks) offer a modular approach to plasmid construction.

Purpose of the Study:

  • To demonstrate the utility of biobricks for creating custom plasmids.
  • To develop a versatile collection of biobricks for diverse genetic engineering applications.

Main Methods:

  • Assembled a library of 52 biobrick parts including multiple cloning sites, protein tags, reporters, and selection markers.
  • Designed parts to maintain reading frame for protein fusions.
  • Constructed model plasmids for protein binding-site motifs, eukaryotic cloning, and chromosomal insertion.

Main Results:

  • Successfully built a cerulean-reporter plasmid in three biobrick iterations for fluorescent protein fusions.
  • Implemented recombinase-mediated DNA insertion (RMDI) for site-directed gene exchange.
  • Demonstrated generation of multiple standardized cell lines from a single recipient line via fluorescent gene exchange.

Conclusions:

  • The biobrick collection provides a versatile platform for constructing custom plasmids.
  • This approach facilitates the creation of standardized cell lines and streamlines genetic engineering.
  • The library can be distributed peer-to-peer and through the Registry of Standard Biological Parts.