Difference between revisions of "Part:BBa K215107"

(Usage and Biology)
(Usage and Biology)
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To test this secretion system's functionality, it was used to secrete [[https://parts.igem.org/Part:BBa_K215010 BBa_K215010]] (GFP fusion protein containing prtB secretion tag at C-terminus).
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To test this secretion system's functionality, it was transformed into cells (strain BL21 lacq) also containing a plasmid with part[[https://parts.igem.org/Part:BBa_K215010 BBa_K215010]] (GFP fusion protein containing prtB secretion tag at C-terminus).
  
 
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Revision as of 02:34, 19 October 2009

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

For full characterization data on this part, please see the [http://2009.igem.org/Team:Washington/Project/Secretion University of Washington 2009 iGEM project page].


To test this secretion system's functionality, it was transformed into cells (strain BL21 lacq) also containing a plasmid with part[BBa_K215010] (GFP fusion protein containing prtB secretion tag at C-terminus).

Based on these results, it has yet to be shown that the Secretion system is functioning. However, this system has been shown to work properly in the literature, and there are many parameters that we have yet to reproduce and optimize. Proteins that have been secreted by this Type I system and other similar ones include GFP, lipase, Trichoderma harzianum endochitinase, trout growth hormone, ompC, and lacZ. This causes us to believe that this part, which has been sequenced verified, should work given a little more tweaking, to see our ideas see Future Directions



Continue to [http://2009.igem.org/Team:Washington/Project/Display Display]

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