Regulatory

Part:BBa_K4628019

Designed by: Linghao Xia   Group: iGEM23_ZJU-China   (2023-10-11)


subPromoter MP

Usage and Biology

The sgRNA promoter for the MP gene is located within the ORF of the 183-kDa protein that is required for TMV replication. The core and the full MP sgRNA promoters were between 235 and 110 and between 295 and 140, respectively, relative to the transcription start site. Computer analysis predicted two dry ring structures (SL1 and SL2) upstream of MP sgRNA transcription initiation sites. Deletion analysis and site-specific mutation showed that the MP sgRNA promoter activity required the SL1 secondary structure, not its sequence, but the deletion of -39nt in the SL2 region increased the accumulation of MP sgRNA fourfold.

This Part is derived from the subgenomic movement protein promoter in the TMV genome, which mediates the transcription of MP.

Because part of the movement protein coding sequence is included in the TMV RNA-dependent RNA polymerase coding sequence, it means that the retention of intact replicases and subgenomic promoters may result in meaningless partial movement protein peptide chains being transcribed and translated, affecting the correct folding and functional expression of the target protein. However, our results successfully prove that it is feasible to express GFP with this promoter without affecting its normal luminescence(Fig.1)

subPromoter MP

Figure 1 subPromoter MP:EGFP in the tobacco leaf

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]


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Categories
Parameters
biologytobacco mosaic virus
chassisNicotiana benthamiana