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Team:Washington/Magnetosomes/Background
From 2011.igem.org
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=== Gibson Assembly Toolkit === | === Gibson Assembly Toolkit === | ||
| - | As an expansion of work started by the [https://2010.igem.org/Team:Washington/Tools_Used/Next-Gen_Cloning 2010 UW IGEM team], this year we developed and submitted a set of plasmid backbones for | + | As an expansion of work started by the [https://2010.igem.org/Team:Washington/Tools_Used/Next-Gen_Cloning 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 [http://dspace.mit.edu/bitstream/handle/1721.1/46747/BBFRFC21.pdf?sequence=1 RFC 21], these "pGA" vectors comprise the [https://2011.igem.org/Team:Washington/Magnetosomes/GibsonVectors Gibson Assembly Toolkit] |
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;'''What's in the Gibson Assembly Toolkit?'''<br/> | ;'''What's in the Gibson Assembly Toolkit?'''<br/> | ||
*Five plasmid backbones | *Five plasmid backbones | ||
| - | * 2 High copy extraction vectors: pGA1A3, pGA1C3 | + | * 2 High copy extraction/cloning vectors: pGA1A3, pGA1C3 |
* 3 low copy assembly vectors: pGA3K3, pGA4A5, pGA4C5 | * 3 low copy assembly vectors: pGA3K3, pGA4A5, pGA4C5 | ||
Revision as of 02:10, 23 September 2011
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 BioBricks that are optimized for Gibson assembly. Based on the bglBrick standard [http://dspace.mit.edu/bitstream/handle/1721.1/46747/BBFRFC21.pdf?sequence=1 RFC 21], these "pGA" vectors comprise the Gibson Assembly Toolkit
- What's in the Gibson Assembly Toolkit?
- Five plasmid backbones
- 2 High copy extraction/cloning 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
