Part:BBa_K4229067
mTurquoise2 with C-terminal SnoopCatcher regulated by tetA/B promotor
This composite BioBrick was used to visualize protein targeting into microcompartments by fluorescent microscopy. The SnoopCatcher is fused to the C-term of mTurquoise2.
We used this fusion protein to test the functionality of our compartments: the minimal wiffleball (BBa_K4229047), full wiffleball (BBa_K4229049), and SPD-5 (BBa_K4229078). All compartmentalization proteins have an N-terminal SpyTag.
A fluorescent signal in the bacteria that expressed either protein could be detected. Upon co-expression with compartmentalization proteins, foci were formed (especially well with the full wiffleball) with both reporter genes:
It was later confirmed that in fact mTurqouise was cached by T1 by a western blot comparing the full wiffleball and the minimal wiffleball.
With that, we can say that our mTurquiose2 with the snoopCatcher is successfully expressed and caught by our compartment.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 59
Illegal PstI site found at 856 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 59
Illegal PstI site found at 856 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 59
Illegal BglII site found at 68 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 59
Illegal PstI site found at 856 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 59
Illegal PstI site found at 856
Illegal AgeI site found at 121
Illegal AgeI site found at 1017 - 1000COMPATIBLE WITH RFC[1000]
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