Device

Part:BBa_K554015

Designed by: UNICAMP EMSE Brazil team   Group: iGEM11_UNICAMP-EMSE_Brazil   (2011-09-26)

flhDC HlyB HlyD TolC device

This part represents the final assembly of the hemolysin secretion system ([http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Project#Device_3:_Secretion_system Device 3, Protein Secretion System]), very important to export the proteins produced inside bacteria and that must act in targets outside (such as IL-12 and IL-10). This device is composed by flhDC promoter linked to Hemolysin B, Hemolysin D, TolC and Terminator.

This is the Secretion device used by [http://2011.igem.org/Team:UNICAMP-EMSE_Brazil UNICAMP-EMSE Brazil team] to export IL-12, since it is regulated by the promoter flhDC. The main purpose of not using constitutive production of this secretion system is to avoid energy waste, then bacteria would only produce these proteins under the specific signal (in this case, presence of Catecholamines or AI-3).

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 1394
    Illegal BglII site found at 2057
    Illegal BamHI site found at 4101
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 324
    Illegal AgeI site found at 3561
    Illegal AgeI site found at 3618
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 3953


Usage and Biology

You can see a representation of this device acting in the schema below: HlyB, HlyD and TolC join together to produce the protein export system shown as a symbolic cilinder in orange.

UNICAMP EMSE secretion device flhdc schema.jpg

Representation of device 3, the protein secretion system, in a Jedi bacteria that contains Device 1 (Adrenaline sensor/IL-12 producer). To export a protein, the bacteria must have the HlyD, HlyB and TolC proteins and the target protein must have a signal sequence (HlyA tail). In this case, the target protein to be secreted is IL-12.

A more realistic schema of ABC transport system is shown below:

UNICAMP EMSE secretion ABC.jpg
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Categories
Parameters
None