Part:BBa_K4990016
MazEF system
To mitigate leakage risks such as engineering bacteria in hypoxic environments like sewers after fecal excretion, we developed a temperature-responsive suicide switch. For our external suicide switch design, we drew on BNU-China 2019 project research, specifically their use of the TA system which piqued our interest. However, it should be noted that according to Fudan's 2020 proposal, the RelE/RelB module used by BNU-China in 2019 has further effects on the human gut. RelE possesses specific activity in eukaryotes and has been observed to cause cell apoptosis in cultured human cell lines. Additionally, RelE can accumulate unexpectedly in bacteria and affect the bacteria intricately involved in the shaping of human gut microbiota. After conducting research, it has been determined that the Type II TA system is the most thoroughly examined system.[1] Additionally, among these systems, the MazE/MazF module is considered to be relatively well-explored. Consequently, we have opted to implement the MazEF module in our suicide switch design. For regulation at the translation level, we have installed a temperature-sensitive RNA switch upstream of the antitoxin mazE. When the engineered bacteria are exposed to a high-temperature environment (37℃ in vivo), the RNA switch is activated, resulting in normal translation of both the antitoxin and toxin. The antitoxin typically hinders the toxin's effects, and the genetically modified bacteria can endure. At low-temperature environment(25℃ in vitro), the RNA switch forms a hairpin structure obstructing the antitoxin's translation and granting way to regular toxin translation and start suicide program.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 137
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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