Difference between revisions of "Part:BBa K3332039"
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'''Fig 1.''' Kill switch of the detection part
 
'''Fig 1.''' Kill switch of the detection part
  
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In this circuit, LacI can repress pTrc-2 promoter and pTrc-2 derivative promoter ，while tetR can repress pLtetO-1 promoter. When ATc exits, it combines with tetR, so that pLtetO-1 promoter can’t be repressed. Then LacI which is controlled by pLtetO-1 can repress the pTrc-2 promoter and pTrc-2 derivative promoter. As a result, mf-lon and MazF can’t be expressed.  
+
In this circuit, LacI can repress pTrc-2 promoter and pTrc-2 derivative promoter ，while tetR can repress pLtetO-1 promoter. When ATc exits, it combines with tetR, so that pLtetO-1 promoter can’t be repressed. Then LacI which is controlled by pLtetO-1 can repress pTrc-2 promoter and pTrc-2 derivative promoter. As a result, mf-lon and mazF can’t be expressed.  
  
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As a kind of bacterial toxin, the expression of MazF often lead to the death of bacteria. So there comes the conclusion that as long as the engineered ''E.coli'' are cultured in the environment with ATc, it won’t be killed by the mazF, but when the ''E.coli'' escape from our detection instrument, the effect can be reversed. That is to say, the ''E.coli'' will be killed by MazF. In the same way, we can see that in the presence of IPTG, MazF can be expressed and kill the''E.coli''.
+
As a kind of bacterial toxin, the expression of mazF often lead to the death of bacteria. So there comes the conclusion that as long as the engineered ''E.coli'' are cultured in the environment with ATc, it won’t be killed by mazF, but when the ''E.coli'' escape from our detection instrument, the effect can be reversed. That is to say, the ''E.coli'' will be killed by MazF. In the same way, we can see that in the presence of IPTG, mazF can be expressed and kill the''E.coli''.
  
 
===Sequence and Features===
 
===Sequence and Features===

Revision as of 18:46, 27 October 2020


tetR

The tetR protein is able to repress pLtetO-1 promoter in the absence of ATc.

Usage and Biology

The tetR protein is used to inhibit pLtetO-1. It is part of the circuit designed to prevent engineered bacteria in the detection instrument from escaping.

Fig 1. Kill switch of the detection part

In this circuit, LacI can repress pTrc-2 promoter and pTrc-2 derivative promoter ，while tetR can repress pLtetO-1 promoter. When ATc exits, it combines with tetR, so that pLtetO-1 promoter can’t be repressed. Then LacI which is controlled by pLtetO-1 can repress pTrc-2 promoter and pTrc-2 derivative promoter. As a result, mf-lon and mazF can’t be expressed.

As a kind of bacterial toxin, the expression of mazF often lead to the death of bacteria. So there comes the conclusion that as long as the engineered E.coli are cultured in the environment with ATc, it won’t be killed by mazF, but when the E.coli escape from our detection instrument, the effect can be reversed. That is to say, the E.coli will be killed by MazF. In the same way, we can see that in the presence of IPTG, mazF can be expressed and kill theE.coli.

Sequence and Features


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]


Reference

[1] Chan CT, Lee JW, Cameron DE, Bashor CJ, Collins JJ. 'Deadman' and 'Passcode' microbial kill switches for bacterial containment. Nat Chem Biol. 2016;12(2):82-86. doi:10.1038/nchembio.1979