Device
p_mles+RBS

Part:BBa_K4137014:Design

Designed by: YA-JHIH KO   Group: iGEM22_GEMS_Taiwan   (2022-10-07)
Revision as of 08:08, 10 October 2022 by Andreahuang (Talk | contribs)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)

CcdA regulatory and expression device


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 419
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We chose to use p_mles, the mleR regulatory site as mleR can be induced by malate acid which surrounds the tree roots. CcdA is the antitoxin of the toxin Ccdb which kills chi18h8 when the probiotic isn’t near the roots of the banana tree. The Myc tag is used for protein quantification and is one of the most commonly used purification tag. The T7 promoter enables over production of mleR. RBS (AAGGAG) has been shown to be efficient in protein expression MleR can be regulated by malate acid, which is presented around the roots. MleR will be further used as a signal for the production of antitoixen CcdA. The 6xHis tag fused to mleR allows the isolation of mleR for quantification and purification.

Figure 2. ccdA-mleR Benchling linear map.

Source

https://www.ncbi.nlm.nih.gov/nuccore/X75982.1 https://www.snapgene.com/resources/plasmid-files/?set=pet_and_duet_vectors_(novagen)&plasmid=pET-28a(%2B) https://www.ncbi.nlm.nih.gov/nuccore/U51588.1 https://www.snapgene.com/resources/plasmid-files/?set=pet_and_duet_vectors_(novagen)&plasmid=pET-28a(%2B) https://www.snapgene.com/resources/plasmid-files/?set=pet_and_duet_vectors_(novagen)&plasmid=pET-28a(%2B) https://journals.asm.org/doi/10.1128/jb.171.6.3108-3114.1989

References