Difference between revisions of "Part:BBa K743012"

Revision as of 16:30, 22 September 2012

OmpA + HIV protease cleavage site - GFP secretion system

Arabinose -> extracellular GFP

Upon addition of arabinose to the medium, the inducible Pbad promoter drives the expression of outer membrane protein A fused to a extracellular linker (ompA brick) followed by a cleavage site recognizable by HIV protease wich is linked to GFP coding sequence.

If HIV protease is present in the medium, it will cut the cleavage site and GFP will be released from the outer membrane

It was used by [http://2012.igem.org/Team:UC_Chile 2012 UC Chile team] to secrete GFP. You can check the results HERE LINK!

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 1205
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 1144
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 979
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI site found at 961
Illegal SapI.rc site found at 1743

@@ Line 4: / Line 4: @@
 '''Arabinose -> extracellular GFP'''
-Upon addition of arabinose to the medium, the inducible Pbad promoter drives the expression of outer membrane protein A  fused to a extracellular linker ([https://parts.igem.org/Part:BBa_K103006 ompA brick]) followed by a cleavage site recognizable by HIV protease wich is linked to GFP coding sequence.
+Upon addition of arabinose to the medium, the inducible Pbad promoter drives the expression of outer membrane protein A  fused to a extracellular linker ([https://parts.igem.org/Part:BBa_K103006 ompA brick]) followed by a cleavage site recognizable by [https://parts.igem.org/Part:BBa_I712667 HIV protease] wich is linked to GFP coding sequence.
 If HIV protease is present in the medium, it will cut the cleavage site and GFP will be released from the outer membrane