Team:TU-Eindhoven/LEC/BioBricks

From 2012.igem.org

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<groupparts>iGEM012 TU-Eindhoven</groupparts>
<groupparts>iGEM012 TU-Eindhoven</groupparts>
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<p>Our iGEM team prepared three different BioBricks: BBa_K881000 (R-GECO), BBa_K881001 (G-GECO), BBa_K881002 (B-GECO). These three GECO proteins, which are obtained from ''Zhao et al. 2011''<html><a href="#ref_zhao" name="text_zhao"><sup>[1]</sup></a></html>, are red, green and blue fluorescent respectively. The coding DNA for these three proteins was ligated into a <partinfo>pSB1C3</partinfo> vector. After culturing, the vectors were restricted with restriction enzymes XbaI en PstI. These enzymes restricted the vector nicely, however, the DNA sequence coding for the fluorescent GECO proteins appears to contain a restriction site for PstI aswell. Therefore, this procedure was repeated with restriction enzymes XbaI and SpeI.
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<p>Our iGEM team prepared three different BioBricks: BBa_K881000 (R-GECO), BBa_K881001 (G-GECO), BBa_K881002 (B-GECO). These three GECO proteins, which are obtained from ''Zhao et al. 2011''<html><a href="#ref_zhao" name="text_zhao"><sup>[1]</sup></a></html>, are red, green and blue fluorescent respectively. The coding DNA for these three proteins was ligated into a <partinfo>pSB1C3</partinfo> vector. After culturing, the vectors were restricted with restriction enzymes XbaI en PstI. These enzymes restricted the vector nicely, however, the DNA sequence coding for the fluorescent GECO proteins appears to contain a restriction site for PstI aswell. Therefore, this procedure was repeated with restriction enzymes XbaI and SpeI.</p>
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<p>Although, these BioBricks are prepared in a more efficient way using our own protocol, the permission we needed to apply the GECO BioBricks to registry library was not easy to obtain. You can read more about this struggle at the 'Considerations' page.</p>
The GECO’s, R-GECO (red), G-GECO (green) and B-GECO (blue) are the biobricks we will send to the registry. Those biobricks can be used in yeast and in E.coli cells. The protocol attached with the biobrick-library didn’t work for us. We used our own restriction and ligation protocols to develop the biobrick.</p>
The GECO’s, R-GECO (red), G-GECO (green) and B-GECO (blue) are the biobricks we will send to the registry. Those biobricks can be used in yeast and in E.coli cells. The protocol attached with the biobrick-library didn’t work for us. We used our own restriction and ligation protocols to develop the biobrick.</p>
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<p>The permission we needed to add the GECO’s to registry library was not easy to get. You can read more about this struggle at the Considerations-page.</p>
 

Revision as of 20:44, 25 September 2012