Difference between revisions of "Part:BBa K4137014:Design"
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The 6xHis tag fused to mleR allows the isolation of mleR for quantification and purification. | The 6xHis tag fused to mleR allows the isolation of mleR for quantification and purification. | ||
− | + | [[File:ccda-mler-benchling.png|750px|thumb|center|Figure 2. ccdA-mleR Benchling linear map.]] | |
===Source=== | ===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.ncbi.nlm.nih.gov/nuccore/U51588.1 | ||
− | https:// | + | https://www.snapgene.com/resources/plasmid-files/?set=pet_and_duet_vectors_(novagen)&plasmid=pET-28a(%2B) |
− | https:// | + | 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=== | ===References=== |
Latest revision as of 08:08, 10 October 2022
CcdA regulatory and expression device
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 419
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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.
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