Difference between revisions of "Part:BBa K208049"

Revision as of 16:06, 22 October 2009

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.

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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 362
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 881
Illegal XhoI site found at 782
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 __NOTOC__
 <partinfo>BBa_K208049 short</partinfo>
-GFP Secretion by TorA
+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.
-<!-- Add more about the biology of this part here
 ===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.
 <!-- -->