Difference between revisions of "Part:BBa K4229067"

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It was later confirmed that in fact mTurqouise2 was cached by T1 by a western blot comparing the full wiffleball and the minimal wiffleball.
 
It was later confirmed that in fact mTurqouise2 was cached by T1 by a western blot comparing the full wiffleball and the minimal wiffleball.
[[File: MTurquoiseWestern.png|800px|thumb|left|: Western Blot of the BMC minimal wiffleball (pT1spysnp) and full wiffleball (pT1spysnpT2T3) + mTurquoise2]]
+
[[File:MTurquoiseWestern.png|800px|thumb|left|: Western Blot of the BMC minimal wiffleball (pT1spysnp) and full wiffleball (pT1spysnpT2T3) + mTurquoise2]]
  
  

Revision as of 07:13, 12 October 2022


mTurquoise2 with C-terminal SnoopCatcher regulated by tetA/B promotor

mTurqouise2 is with mVenus2(BBa_K4229066) one of our reporter genes, with which we tested the building and catching of the minimal(BBa_K4229047) and full wiffleball(BBa_K4229049) and SPD5 (BBa_K4229078).

The snoop/spyTag snoop/spyCatcher was tested with those two reporter genes. You can see a fluorescent signal in the bacteria that expressed either of those two. You can see foci (especially well with the full wiffleball) for both of those reporter genes:

Fluorescent microscopy of T1 catching the mVenus2 and mturquoise2 when the minimal or full BMC construct is expressed: A Controls for induction; B T1 with and without the Spy/Snp tags; scalebar 5µm





























It was later confirmed that in fact mTurqouise2 was cached by T1 by a western blot comparing the full wiffleball and the minimal wiffleball.

: Western Blot of the BMC minimal wiffleball (pT1spysnp) and full wiffleball (pT1spysnpT2T3) + mTurquoise2















With that, we can say that our mTurquiose2 with the snoopCatcher is successfully expressed and caught by our compartment.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 59
    Illegal PstI site found at 856
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 59
    Illegal PstI site found at 856
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 59
    Illegal BglII site found at 68
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 59
    Illegal PstI site found at 856
  • 25
    INCOMPATIBLE 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
  • 1000
    COMPATIBLE WITH RFC[1000]