Difference between revisions of "Part:BBa K174011:Design"
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===Design Notes===
 
===Design Notes===
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In ''B. subtilis'', out of five kinases only KinA, KinB and KinC kinases are effective to trigger the sporulation. KinD and KinE do not phosphorylate Spo0F, hence do not effect the sporulation. We chose KinA for our device.
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In ''B. subtilis'', out of the five kinases KinA-E only KinA, KinB and KinC kinases trigger sporulation. KinD and KinE do not phosphorylate Spo0F, hence do not effect sporulation. We therefore chose to overexpress the ''kinA'' gene for our device.
  
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As our whole system grew, it was not possible to test everything. Rather than LacI which can be derepressed by IPTG, we had another transcription factor that represses the device's promoter. Instead, we designed our promoter with flanking sites to be induced by IPTG and unit test it. The idea was to construct everything and plug them all together after we test each component.
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This part has received basic characterisation.
  
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pSpac was included for testing purposes. However we designed our promoter with flanking sites to allow different inducible promoters to be used.
  
 
===Source===
 
===Source===
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===References===
 
===References===
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#Veening, J.-W., W. K. Smits, et al. (2008). "Bistability, Epigenetics, and Bet-Hedging in Bacteria." Annual Review of Microbiology 62(1): 193-210.
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#Fujita, M. and R. Losick (2005). "Evidence that entry into sporulation in ''Bacillus subtilis'' is governed by a gradual increase in the level and activity of the master regulator Spo0A." Genes & Development 19(18): 2236-2244.

Latest revision as of 21:17, 21 October 2009

IPTG inducable kinA Bacillus sporulation trigger


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

In B. subtilis, out of the five kinases KinA-E only KinA, KinB and KinC kinases trigger sporulation. KinD and KinE do not phosphorylate Spo0F, hence do not effect sporulation. We therefore chose to overexpress the kinA gene for our device.

This part has received basic characterisation.

pSpac was included for testing purposes. However we designed our promoter with flanking sites to allow different inducible promoters to be used.

Source

Synthesised.

References

  1. Veening, J.-W., W. K. Smits, et al. (2008). "Bistability, Epigenetics, and Bet-Hedging in Bacteria." Annual Review of Microbiology 62(1): 193-210.
  2. Fujita, M. and R. Losick (2005). "Evidence that entry into sporulation in Bacillus subtilis is governed by a gradual increase in the level and activity of the master regulator Spo0A." Genes & Development 19(18): 2236-2244.