Coding

Part:BBa_K729004:Experience

Designed by: Bouran Sohrabi   Group: iGEM12_University_College_London   (2012-06-30)
Revision as of 18:15, 21 October 2014 by Ddelatorre (Talk | contribs) (→‎Applications of BBa_K729004)


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Applications of BBa_K729004

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Testing the compatibility of the nuclease with Azo-dyes


The <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K729004">BBa_K729004</a> periplasmic nuclease that UCL iGEM 2012 <a href="http://2012.igem.org/Team:University_College_London/Module_6/Results">submitted and characterised</a> was able to demonstrate positive DNase agar assays. This assay involves washing DNA-containing agar with HCl after streaking bacteria. After this wash, a ‘halo’ surrounding the streaked bacteria indicates that extracellularly secreted DNase digested the DNA surrounding the colonies within the agar. In order to further characterise this BioBrick and incorporate it into our project as a biosafety method of minimising the transfer of extracellular DNA, we decided to test whether BBa_K729004 would function in the presence of Azo-Dyes.




<img src="Nuclease_fig1.jpg"width="80%">


Figure 1 - <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K729004">BBa_K729004</a> BBa_K729004 periplasmic nuclease enzyme shows functionality in the presence of multiple Azo-dyes.  Figure showing that the BBa_K729004 periplasmic nuclease enzyme is still able to digest the surrounding DNA in the DNase agar. Figure 1a and 1b demonstrate the presence of halos around colonies on plates with and without Acid Orange 7 (AO7) azo-dye. Figure 1c and 1d demonstrate the presence of halos around colonies on plates with and without Reactive Black 5 (RB5). All azo-dye agar plates were made with a 1:500 dilution of 0.5mgml-1 dye.



Since the azo-dye degradation would be taking place in industrial environments, being the products of the bioreaction at one point dumped into other channels of water-treatment or into the environment, it is essential that there are certain barriers that stop the DNA from our genetically modified organisms from potentially being transferred into wild-type strains. On a basic first-level defense, this assay suggests that horizontal gene transfer could be inhibited in an azo-dye contaminated environment by<a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K729004">BBa_K729004</a> , as it has proven effective in degrading extracellular bacterial DNA.


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By UCL 2014

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