Team:Washington/Magnetosomes/Background

From 2011.igem.org

(Difference between revisions)
(Gibson Assembly Toolkit)
(Gibson Assembly Toolkit)
Line 10: Line 10:
=== 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 cloning and expression of BioBricks that are optimized for Gibson assembly. We call these pGA vectors - they comprise the [https://2011.igem.org/Team:Washington/Magnetosomes/GibsonVectors 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 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]  
Line 17: Line 17:
;'''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


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

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


Igem2011 GibsonToolkit.png
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.


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