Difference between revisions of "Part:BBa K4229066"

Revision as of 07:00, 12 October 2022

mVenus with N-terminal spyCatcher regulated by tetA/B promotor

mVenus2 is with mTurqouise2(BBa_K4229067) 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 mVenus was cached by T1 by a western blot comparing the full wiffleball with and without the necessary tags.

Western Blot comparison of the BMC minimal wiffleball with and w/o tags (pHT1) + mVenus2

With that, we can say that our mVenus two with the spyCatcher is successfully expressed and caught by our compartment.

Sequence and Features