Difference between revisions of "Part:BBa K137118"

 
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<partinfo>BBa_K137118 short</partinfo>
 
<partinfo>BBa_K137118 short</partinfo>
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AHL inducible promoter with a medium strength RBS is used to induce rcsA. rcsA leads to induction of lambda phage lysogens.  
 
AHL inducible promoter with a medium strength RBS is used to induce rcsA. rcsA leads to induction of lambda phage lysogens.  
  
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===Usage and Biology===
 
===Usage and Biology===
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The phage λcam was used to establish lysogens of ''E. coli'' strain DH10B carrying rcsA induction devices in pSB1A2. Using the Caltech iGEM [http://2008.igem.org/Team:Caltech/Protocols/rcsA_Lysogen_Induction rcsA lysogen induction protocol] and then [http://2008.igem.org/Team:Caltech/Protocols/Titering titering] the resulting phage on wild-type ''E. coli'', the series of devices S04080 (no promoter), K137129 (weak RBS B0033), K137118 (medium RBS B0032), K137114 (strong RBS B0034), and K137117 (strong RBS B0034 plus constitutive cI expression) produced the following concentrations of plaque forming units (pfu/mL).
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[[Image:RcsA.gif|700px]]
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Increasing the RBS strength increased the background lysogen induction without changing the dynamic range (ratio of pfu/mL induced vs. uninduced). Adding constitutive cI expression increased the dynamic range roughly 3-fold, lowering background leakiness without significantly lowering the induced expression.
  
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K137118 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K137118 SequenceAndFeatures</partinfo>

Revision as of 01:17, 22 October 2008

AHL induced production of rcsA; medium strength

AHL inducible promoter with a medium strength RBS is used to induce rcsA. rcsA leads to induction of lambda phage lysogens.

Usage and Biology

The phage λcam was used to establish lysogens of E. coli strain DH10B carrying rcsA induction devices in pSB1A2. Using the Caltech iGEM [http://2008.igem.org/Team:Caltech/Protocols/rcsA_Lysogen_Induction rcsA lysogen induction protocol] and then [http://2008.igem.org/Team:Caltech/Protocols/Titering titering] the resulting phage on wild-type E. coli, the series of devices S04080 (no promoter), K137129 (weak RBS B0033), K137118 (medium RBS B0032), K137114 (strong RBS B0034), and K137117 (strong RBS B0034 plus constitutive cI expression) produced the following concentrations of plaque forming units (pfu/mL).

RcsA.gif

Increasing the RBS strength increased the background lysogen induction without changing the dynamic range (ratio of pfu/mL induced vs. uninduced). Adding constitutive cI expression increased the dynamic range roughly 3-fold, lowering background leakiness without significantly lowering the induced expression.

Sequence and Features


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1005