Composite

Part:BBa_K3215013

Designed by: Daniel Oppermann Peixoto   Group: iGEM19_UFRGS_Brazil   (2019-09-08)
Revision as of 16:03, 20 October 2019 by Danielopeixoto (Talk | contribs)


Tse2 Repressed by Arabinose Kill Switch

This kill switch was created to enable its user to reduce an organism's growth rate in a medium without arabinose. It is consisted of parts BBa_K259007, BBa_K327018, BBa_B0015, BBa_R0051, BBa_J61100 and BBa_K314200.

Usage and Biology

Considering a system where the survival of an organism is inadequate, this BioBrick displays a way of handling the issue. When this mechanism is present, the presence of L-(+)-Arabinose will enhance the transcription of cI, which will repress the expression of the Tse2 toxin, enabling the survival of the organism. On the other hand, when the former is absent, the transctiption of cI will be inhibited, which in turn will activate the expression of Tse2 toxin, repressing the survival rate of those organisms. For our purposes, this explanation lacks one major detail: cAMP. When glucose is absent in the media, the levels of cAMP will rise, and that can disrupt the balance of the system, knowing that cAMP will bind to CRP, enabling the transcription of cI, interfering in the Arabinose regulation.

2019 UFRGS Brazil Tse2 scheme.png

Fig. 1. BBa_K3215013 scheme.


Characterization

To characterize and validate our part, two different approaches were made:

Dry lab experiment validation

Our team was able to create a mathematical model that correlates the effects of Arabinose concentration with the levels of cI and Tse2, speculating cell survival and variations of the efficiency of the model in different concentrations of extracellular glucose. To see all the details of our model, check section 2 from our modelling page (<a href="https://2019.igem.org/Team:UFRGS_Brazil">Team 2019 UFRGS_Brazil</a>)

Wet lab experiment validation

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 294
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1532
    Illegal NgoMIV site found at 1589
    Illegal NgoMIV site found at 1714
    Illegal AgeI site found at 125
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 107


[edit]
Categories
//biosafety/kill_switch
Parameters
None