Team:Groningen/Project/Backbone

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To get our developed biobricks into b. subtilis, a backbone was developed. The biobrick compatible backbone developed by the iGEM team of Munich 2012 (Insert exact name)  was used as an template but is improved (insert link to part registry) . A hyspank promoter was added to the backbone. The multiple cloning site is used to enter our biobricks into the system. For the use of b. subtilis. the amyE sites are used to cut it
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To transform <i>B. subtilis</i> with our developed biobricks we have improved the <i>amyE</i> intergrational backbone from Munich's iGEM team 2012 (<a href="http://parts.igem.org/Part:BBa_K823023">BBa_K823023</a>) that had no inducible promoter. We've added the HyperSpank IPTG inducible promoter from Rudner's Lab 2004. This promoter is placed right in front of the prefix, so any biobrick can be inserted in this backbone (<a href="http://parts.igem.org/Part:BBa_K1085014">BBa_K1085014</a>).
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We used the empty backbone for integration into <i>Bacillus subtilis amyE</i> locus from Munich 2012 (BBa_K823023) that had no promoter. We've added the Hy_Spank IPTG inducible promoter from Rudner's Lab 2004. The Hy_Spank promoter is right in front of the prefix so any biobrick can be inserted in this backbone.  
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Revision as of 19:35, 30 September 2013

Backbone

To transform B. subtilis with our developed biobricks we have improved the amyE intergrational backbone from Munich's iGEM team 2012 (BBa_K823023) that had no inducible promoter. We've added the HyperSpank IPTG inducible promoter from Rudner's Lab 2004. This promoter is placed right in front of the prefix, so any biobrick can be inserted in this backbone (BBa_K1085014).

Figure 1: Hy_Spank promoter

Figure 2:

Promoter activity


Figure 3: The intensity in of ~40 cells in the GFP-channel, were analyzed from two pictures.
The average intensity (AU) from the cells are plotted above with the standard deviation.

GFPmg and GFP (BBa_E0840)


Figure 4:



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