Difference between revisions of "Part:BBa K3132101"
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<partinfo>BBa_K3132101 short</partinfo> | <partinfo>BBa_K3132101 short</partinfo> | ||
| − | + | 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.
References
[1] M. Fussenegger et al., Streptogramin-based gene regulation systems for mammalian cells. Nature biotechnology 18, 1203--1208 (2000).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]