Part:BBa_K554013
SoxS 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 SoxS 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-10 (or GFP in the testing devices) ([http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Project#Device_2:_NO_sensor.2FIL-10_producer NO sensor device/ IL-10 producer] (from "Device 2", which senses NO levels and responds by producing and secreting IL-10), since it is regulated by the promoter SoxS. 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 Nitric Oxide or Superoxide).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1371
Illegal BglII site found at 2034
Illegal BamHI site found at 4078 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 301
Illegal AgeI site found at 3538
Illegal AgeI site found at 3595 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 3930
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. In our experiments, we used [http://en.wikipedia.org/wiki/Paraquat Paraquat] in order to induce oxidative stress in bacteria, which causes a increase of Nitric Oxide (NO).
Representation of device 3, the protein secretion system coupled to Sox regulated GPF production device (Part BBa_K554012). 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), as shown here in a light red oval. In this case, the target protein to be secreted is GFP.
A more realistic schema of ABC transport system is shown below:
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