Part:BBa_K4656010:Design
PRM-CI857-PR-phIF-MazE-PphIF-MazF
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1625
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
In summer, the temperature in some areas can reach more than 37℃, resulting in engineered bacteria in these places can survive even if they leave the human body. So there is still a risk of damaging the environment after being discharged from the human body. This point needs to be considered carefully.
Source
The temperator-sensitive lambda CI repressor protein has a CI857 mutation, and when the temperature is reduced to 30℃, CI857 converges to synthesize a dimer, binds to the promoter PR and inhibits it, resulting in suppression of downstream phIF expression. Because the phIF expression product can inhibit the promoter PphIF and inhibit the expression of the toxin MazF downstream of it. Therefore, when the temperature decreased, the inhibition ability of phIF to promoter PphIF was weakened or even lost, and MazF expressed a large number of toxic proteins, resulting in the death of E. coli. However, when the temperature increased to 37℃~42℃, CI857 repressor protein denaturated, CI repressor could no longer bind to promoter PR and play a repressor role, and a large number of downstream phIF protein inhibited promoter PphIF. Therefore, the expression of toxin MazF in the pathway was inhibited, and the MazE antitoxin protein at the end of the gene route was expressed instead. Eventually the E. coli survived.