Part:BBa_K215107
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 used to secrete [BBa_K215010] (GFP fusion protein containing prtB secretion tag at C-terminus).
Tagged GFP in Media The red data set corresponds to cells containing this secretion system plasmid and part BBa_K215011 (fusion OpdA with prtB tag), the blue data set corresponds to cells containing a promoter-less version of the secretion system and part BBa_K215010 (fusion GFP with prtB tag), and the green data set corresponds to cells containing this secretion system and part BBa_K215010. 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 is rather an artifact.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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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