Difference between revisions of "Part:BBa K1638032"
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This part contains the T18 domain of the adenylate cyclase <i>cyaA</i> from <i>Bordetella pertussis</i>. This part is intented to be used in the bacterial two-hybrid system. This system is based on the reconstitution of the adenylate cyclase. When protein-coding genes is suffixed to the T25 and T18 domains of <i>cyaA</i>, one can study the interaction of these two proteins. If the conjugated proteins interacts, T25 and T18 will be brought into close proximity. This will enable the two catalytic domains T18 and T25 to synthesize cyclic adenosinemonophosphate (cAMP).
 
This part contains the T18 domain of the adenylate cyclase <i>cyaA</i> from <i>Bordetella pertussis</i>. This part is intented to be used in the bacterial two-hybrid system. This system is based on the reconstitution of the adenylate cyclase. When protein-coding genes is suffixed to the T25 and T18 domains of <i>cyaA</i>, one can study the interaction of these two proteins. If the conjugated proteins interacts, T25 and T18 will be brought into close proximity. This will enable the two catalytic domains T18 and T25 to synthesize cyclic adenosinemonophosphate (cAMP).
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The rise in cAMP can trigger the expression of genes by using a cAMP-induced promotor that induce the transcription of red fluorescent protein (RFP). For this reason this part also contains a mRFP generator controlled by PcstA <partinfo>BBa_K861173</partinfo>. The presence of red-fluorescent cells can in turn be used to verify protein-protein interactions.
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The rise in cAMP can trigger the expression of genes by using a cAMP-induced promotor that induce the transcription of red fluorescent protein (RFP). For this reason this part also contains a mRFP generator controlled by PcstA <partinfo>BBa_K861173</partinfo>. The presence of red-fluorescent cells can in turn be used to verify protein-protein interactions [1].
  
 
See <partinfo>BBa_K1638033</partinfo> for the T25 domain.
 
See <partinfo>BBa_K1638033</partinfo> for the T25 domain.
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<partinfo>BBa_K1638032 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1638032 SequenceAndFeatures</partinfo>
  
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===References===
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[1] Karimova G, Pidoux J, Ullmann A, Ladant D. A bacterial two-hybrid system based on a reconstituted signal transduction pathway. Proceedings of the National Academy of Sciences of the United States of America. 1998;95(10):5752-6.
 
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===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K1638032 parameters</partinfo>
 
<partinfo>BBa_K1638032 parameters</partinfo>
 
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Revision as of 00:36, 18 September 2015

T18 domain of cyaA with cAMP-induced RFP generator

This part contains the T18 domain of the adenylate cyclase cyaA from Bordetella pertussis. This part is intented to be used in the bacterial two-hybrid system. This system is based on the reconstitution of the adenylate cyclase. When protein-coding genes is suffixed to the T25 and T18 domains of cyaA, one can study the interaction of these two proteins. If the conjugated proteins interacts, T25 and T18 will be brought into close proximity. This will enable the two catalytic domains T18 and T25 to synthesize cyclic adenosinemonophosphate (cAMP). The rise in cAMP can trigger the expression of genes by using a cAMP-induced promotor that induce the transcription of red fluorescent protein (RFP). For this reason this part also contains a mRFP generator controlled by PcstA BBa_K861173. The presence of red-fluorescent cells can in turn be used to verify protein-protein interactions [1].

See BBa_K1638033 for the T25 domain.

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 1719
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1260
    Illegal NgoMIV site found at 1670
    Illegal AgeI site found at 712
    Illegal AgeI site found at 824
    Illegal AgeI site found at 1476
  • 1000
    COMPATIBLE WITH RFC[1000]

References

[1] Karimova G, Pidoux J, Ullmann A, Ladant D. A bacterial two-hybrid system based on a reconstituted signal transduction pathway. Proceedings of the National Academy of Sciences of the United States of America. 1998;95(10):5752-6.