Difference between revisions of "Part:BBa K3132101"

 
 
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<partinfo>BBa_K3132101 short</partinfo>
 
<partinfo>BBa_K3132101 short</partinfo>
  
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This part (8*PIP MinimalCPIP_MinimalCMVMV ) is one of our synthetic promoters (SynPro) based on minimalCMV promoter (BBa_K3132003) and eight repeats of PIP binding sites. We inset the 8*PIP 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 PIP_DBD_(G4S) linker_NLS_VP64. The PIP-DBD can bind to the 8*PIP binding sites specifically and the VP64 domain can initiate transcription by activating the MinimalCMV 8*PIP promoter. The corresponding SynTF of PIP_minimalCMV is PIP-VP64 (BBa_K3132004).
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This part (8*PIP MinimalCMV ) is one of our synthetic promoters (SynPro) based on minimalCMV promoter  (<partinfo>BBa_K3132004</partinfo>)  and eight repeats of PIP binding sites. We inset the 8*PIP 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 PIP_DBD_(G4S) linker_NLS_VP64. The PIP-DBD can bind to the 8*PIP binding sites specifically[[#References|[1]]] and the VP64 domain can initiate transcription by activating the MinimalCMV 8*PIP promoter. The corresponding SynTF of PIP_minimalCMV is PIP-VP64 (<partinfo>BBa_K3132003</partinfo>).
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==Characterization of PIP_MinimalCMV Promotor: ==
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We used fluorescence protein mCherry to verify the binding of PIP-DBD to 8*PIR(PIP binding sites), and the results are as follows. We inset mCherry behind 8*PIR and in general the fluorescence intensity is very low; when PIP is added, the fluorescence intensity significantly increased which verifies the binding of PIP-DBD to 8*PIR.
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[[File:T--SMMU-China--UAS_MinimalCMV_Promotor.png ‎| 600px|thumb|center|]]
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==References==
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<p>[1] M. Fussenegger et al., Streptogramin-based gene regulation systems for mammalian cells. Nature biotechnology 18, 1203--1208 (2000).</p>
  
 
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Latest revision as of 15:55, 21 October 2019


PIP_MinimalCMV Promotor

This part (8*PIP MinimalCMV ) is one of our synthetic promoters (SynPro) based on minimalCMV promoter  (BBa_K3132004)  and eight repeats of PIP binding sites. We inset the 8*PIP 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 PIP_DBD_(G4S) linker_NLS_VP64. The PIP-DBD can bind to the 8*PIP binding sites specifically[1] and the VP64 domain can initiate transcription by activating the MinimalCMV 8*PIP promoter. The corresponding SynTF of PIP_minimalCMV is PIP-VP64 (BBa_K3132003).

Characterization of PIP_MinimalCMV Promotor:

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

T--SMMU-China--UAS MinimalCMV Promotor.png

References

[1] M. Fussenegger et al., Streptogramin-based gene regulation systems for mammalian cells. Nature biotechnology 18, 1203--1208 (2000).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]