Team:Washington/Magnetosomes/Background

From 2011.igem.org

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

As an expansion of work started by the 2010 UW IGEM team, this year we developed and submitted a set of plasmid backbones for cloning and expression of BioBricks that are optimized for Gibson assembly. We call these pGA vectors - they comprise the Gibson Assembly Toolkit

What's in the Gibson Assembly Toolkit?

• Five plasmid backbones
• 2 High copy extraction vectors: pGA1A3, pGA1C3
• 3 low copy assembly vectors: pGA3K3, 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.

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