Composite

Part:BBa_K215107

Designed by: Josef Dunbar   Group: iGEM09_Washington   (2009-09-30)
Revision as of 01:51, 19 October 2009 by Jeff (Talk | contribs) (→‎Usage and Biology)

Extracellular Secretion System

This part is made of 3 genes: prtD, prtE, and prtF, that constitute a type I Erwinia chrysanthemi secretion system. The operon is expressed from a strong constitutive promoter, BBa_J23100, and has the translational terminator BBa_B0014. In PSB3T5.

Used to secrete proteins containing prtB C-terminal tag. The prtB C-terminal tag is built into the protein generator [BBa_K215002]. Any protein of interest can be inserted into the protein generator and then secreted when used in conjunction with this secretion system.


Usage and Biology

To test this secretion system's functionality, it was used to secrete [BBa_K215010] (GFP fusion protein containing prtB secretion tag at C-terminus).

  • Tagged GFP in Media As the plot shows, a substantial amount of the tagged GFP was released into the media even when the secretion construct did not have a promoter. This result suggests that the tagged GFP in the media was not the result of the secretion, but rather an artifact.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1487
    Illegal BsaI.rc site found at 1133
    Illegal BsaI.rc site found at 3737


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Categories
Parameters
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