Team:Washington/Magnetosomes/Background

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iGEM Toolkits: Background


As with the expansion of the iGEM competition, many iGEM teams have started to investigate the possibility of working with large-scale genomes. Large-scale gene manipulation often requires the use of tools which allow multiple gene inserts as to bring the cloning project from single gene level to a multiple gene level. However, the current BioBrick standard vectors available through iGEM are not designed for multiple-insert cloning. Therefore, the UW iGEM team decided to research methods to improve cloning efficiency and as a result, two "toolkits" were submitted to the registry.



Gibson Assembly Toolkit

To expand on work started by the 2010 UW IGEM team, this year we developed and submitted a set of plasmid backbones for BioBricks that are optimized for Gibson assembly. Based on the bglBrick standard RFC 21, these "pGA" vectors comprise the Gibson Assembly Toolkit. These vectors have much higher cloning efficiencies than the equivalent pSB vector and are fully compliant with BioBrick RFC 57 developed by the 2010 Cambridge iGEM team.


Igem2011 GibsonToolkit.png

What's in the Gibson Assembly Toolkit?
  • Five plasmid backbones
  • 2 High copy vectors for gene extraction and cloning
        • pGA1A3, pGA1C3
  • 1 medium copy expression vector
        • pGA3K3
  • 2 low copy expression vectors
        • pGA4A5, pGA4C5








Magnetosome Toolkit

In addition, we were also ambitious about assembling a large gene-construct of over 16 kb. Therefore, utilizing our pGA vectors and Gibson cloning methods, the Magnetosome Toolkit was developed with the goal to build magnetic E.Coli; a novel characteristic expressed solely by magnetotactic bacteria, such as Magnetospirillum magneticum strain AMB-1.


Igem2011 MagnetToolkit.png

What’s in the Magnetosome Toolkit?

  • A set of the 18 essential genes for the various steps of magnetosome formation.
  • Our favorite genes in pGA vectors
  • A table compiling individual gene functions from our literature search