Part:BBa_K208049
Lac Promoter/RBS/TorA/GFP/Term
This composite part was designed for the export of GFP out of the cytoplasm of E. coli cells. The TorA signal peptide was used, which targets the GFP protein to the TAT translocation pathway (see the TorA page for more details). Both the GFP and TorA parts are Silver-fusion compatible, which means that the TorA part will be attached to GFP following translation. This particular part appears to be functional. TOP10 E. coli cells glowed brightly green when exposed to UV light. Additionally, in several colonies on plates, there appeared to be higher concentrations of GFP in different parts of the cell, which was not observed in the GFP composite part that did not have a signal peptide attached. More characterization is required, but the part is functional.
NOTE: The scar between the signal peptide and the protein is not presented correctly in the sequence below. The scar is the Silver-fusion scar (ACTAGA), which allows for in-frame fusion. Additionally, the scars before the signal peptide and after the protein are affected by these parts each having the Silver-fusion prefix and suffix.
Usage and Biology
Green fluorescent protein (GFP) translocation, or export out of the cytoplasm, has been well documented (Barrett, 2003; Santini, 2001; Thomas, 2001). Due to its ease of detection, studying GFP in parallel with other recombinant protein secretion mechanisms could provide a framework for determining the functionality of the systems.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 362
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 881
Illegal XhoI site found at 782 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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