Difference between revisions of "Part:BBa K818400"

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This is a coding construct for ''B. subtilis'' to produce AmilCP pigment in response to rotten meat volatiles in the environment. AmilCP is a blue pigment designed by iGEM Uppsala 2011 and the regulator, sboA, is designed by iGEM Groningen 2012.
 
This is a coding construct for ''B. subtilis'' to produce AmilCP pigment in response to rotten meat volatiles in the environment. AmilCP is a blue pigment designed by iGEM Uppsala 2011 and the regulator, sboA, is designed by iGEM Groningen 2012.
  
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===Usage and Biology===
 
===Usage and Biology===
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'''Expression in <i>E. coli</i>'''
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SboA-AmilCP is strongly expressed in <i>E. coli</i>, on plate and in liquid culture, at normal growth conditions. On plate, the blue-purple colour is less visible compared to the cell pellet in liquid culture.
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'''Expression in <i>B. subtilis</i>'''
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SboA-AmilGFP was shown to be very weakly expressed in <i>Bacillus subtilis</i> on LB plate without rotten meat induction (faint color formation after 4 days). This is probably due to the leakiness of the promoter.
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We (iGEM Groningen 2012) tested the expression of SboA-AmilCP in <i>Bacillus subtilis</i> subjected to volatiles from spoiled meat using the same setup as we used to test the sticker design (see our sticker page at http://2012.igem.org/Team:Groningen/Sticker). After this setup, a clear blue-purple color was visible.
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Latest revision as of 02:44, 27 October 2012

AmilCP with sboA regulator

This is a coding construct for B. subtilis to produce AmilCP pigment in response to rotten meat volatiles in the environment. AmilCP is a blue pigment designed by iGEM Uppsala 2011 and the regulator, sboA, is designed by iGEM Groningen 2012.

Usage and Biology

Expression in E. coli SboA-AmilCP is strongly expressed in E. coli, on plate and in liquid culture, at normal growth conditions. On plate, the blue-purple colour is less visible compared to the cell pellet in liquid culture.

Expression in B. subtilis

SboA-AmilGFP was shown to be very weakly expressed in Bacillus subtilis on LB plate without rotten meat induction (faint color formation after 4 days). This is probably due to the leakiness of the promoter.

We (iGEM Groningen 2012) tested the expression of SboA-AmilCP in Bacillus subtilis subjected to volatiles from spoiled meat using the same setup as we used to test the sticker design (see our sticker page at http://2012.igem.org/Team:Groningen/Sticker). After this setup, a clear blue-purple color was visible.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 207
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]