Difference between revisions of "Part:BBa K4137012:Design"
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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 the most commonly used purification his-tag.
 
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 the most commonly used purification his-tag.
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===Characterization===
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Cell harvest for ccdA was performed at 3h. We heated the mixture at 95˚C for 10 minutes, centrifuging at 4˚C, 16000g to extract the supernatant. An additional 95˚C heating and 100 µl PBS were added to eliminate clumping of pellets.
  
 
===Source===
 
===Source===

Revision as of 07:52, 10 October 2022

CcdA antitoxin secretion pathway


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]


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 the most commonly used purification his-tag.

Characterization

Cell harvest for ccdA was performed at 3h. We heated the mixture at 95˚C for 10 minutes, centrifuging at 4˚C, 16000g to extract the supernatant. An additional 95˚C heating and 100 µl PBS were added to eliminate clumping of pellets.

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

References