Difference between revisions of "Part:BBa K3132103"

Latest revision as of 15:57, 21 October 2019

ZF43-8_MinimalCMV Promotor

This part (8* ZF_43-8 MinimalCMV) is one of our synthetic promoters (SynPro) based on minimalCMV promoter (BBa_K3132004) and eight repeats of Zinc Finger_43-8 binding sites. We inset the 8* ZF_43-8 binding sites before the minimalCMV promoter. MinimalCMV promoter is a promoter from the vector pcDNA3.1 with very strong transcriptional activity. In this way, we can use our synthetic transcription factors (SynTFs) which is a fusing protein ZF_43-8 _DBD_(G4S) linker_NLS_VP64. The ZF_43-8-DBD can bind to the 8* ZF_43-8 binding sites specifically[1] and the VP64 domain can initiate transcription by activating the MinimalCMV 8*PIP promoter. The corresponding SynTF of ZF_43-8_minimalCMV is ZF43-8-VP64 (BBa_K3132001).

Characterization of ZF43-8_MinimalCMV Promotor:

We used fluorescence protein mCherry to verify the binding of ZF_43-8-DBD to 8* ZF_43-8(Zinc Finger_43-8 binding sites), and the results are as follows. We inset mCherry behind 8* ZF_43-8 and in general the fluorescence intensity is very low; when ZF_43-8 is added, the fluorescence intensity significantly increased which verifies the binding of ZF_43-8-DBD to 8* ZF_43-8.

References

[1] J. J. Lohmueller, T. Z. Armel, P. A. Silver, A tunable zinc finger-based framework for Boolean logic computation in mammalian cells. Nucleic Acids Research 40, 5180--5187 (2012).

Sequence and Features