Difference between revisions of "Part:BBa K3487005"
 
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<partinfo>BBa_K3487005 short</partinfo>
 
<partinfo>BBa_K3487005 short</partinfo>
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===Description===
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RalR acts as a non-specific endonuclease that can cleave methylated and unmethylated DNA, then leading to cell death. As a novel type I TA system, RalR is the first toxin that functions as an endonuclease in <i>E. coli</i>.  Y.X. Guo,et al found that RalR in <i>E. coli</i> functions differently, acting as a toxin by cleaving DNA, and that it belongs to a type I toxin–antitoxin system. In our project, we used RalR as a toxin protein to cleave the DNA of host cell to avoid the bacterial escape.In our project,we used RalR as a toxin protein to  cleave the DNA of host cell  to avoid the bacterial escape. We test the function of this toxin protein in <i>E. coli</i>. The result is as follow.
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This is a new part in the registry. After being expressed, RalR acts as a non-specific endonuclease that can cleave methylated and unmethylated DNA, leading to cell death. We put it as a poisonous protein into the suicide switch and introduce all the strains we use to prevent organisms from escaping.
 
  
 
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==2020 SZPT-CHINA==
 
==2020 SZPT-CHINA==
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===Result===
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We streaked the engineered strain into a 1Mm IPTG plate for overnight culture, and observed that the engineered strain containing the RalR gene did not grow on the plate.
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The expression of RalR was good under the positive and negative controls, and the colony situation in the corresponding area was significantly less than that of the empty vector control group.  The result is as follow.
  
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===Description===
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[[File:T--SZPT-China--RalR.png|center|500px|]]
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After being expressed, RalR acts as a non-specific endonuclease that can cleave methylated and unmethylated DNA, leading to cell death. We put it as a poisonous protein into the suicide switch and introduce all the strains we use to prevent organisms from escaping.
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===Characterization===
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It is crucial to verify that the new part works in the strain. We still don’t know how much of the endonuclease concentration can kill the chassis cells, which determines whether the suicide switch can work. Therefore, we set up a solid plate medium to detect the expression of endonuclease and check our operating procedures.
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===Result===
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The expression of RalR was good under the positive and negative controls, and the colony situation in the corresponding area was significantly less than that of the empty vector control group.
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[[File:T--SZPT-China--RalR.png|center|200px]]
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<center>pET28a(+), vector positive control ddH2O, negative control RalR, Non-specific endonuclease</center>
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===References===
 
===References===
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Guo Y, Quiroga C, Chen Q, et al. RalR (a DNase) and RalA (a small RNA) form a type I toxin-antitoxin system in Escherichia coli. Nucleic Acids Res. 2014;42(10):6448-6462.
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Guo Y, Quiroga C, Chen Q, et al. RalR (a DNase) and RalA (a small RNA) form a type I toxin-antitoxin system in <i>Escherichia coli</i>. Nucleic Acids Res. 2014;42(10):6448-6462.

Latest revision as of 14:52, 10 October 2021


Non-specific endonuclease

Description

RalR acts as a non-specific endonuclease that can cleave methylated and unmethylated DNA, then leading to cell death. As a novel type I TA system, RalR is the first toxin that functions as an endonuclease in E. coli. Y.X. Guo,et al found that RalR in E. coli functions differently, acting as a toxin by cleaving DNA, and that it belongs to a type I toxin–antitoxin system. In our project, we used RalR as a toxin protein to cleave the DNA of host cell to avoid the bacterial escape.In our project,we used RalR as a toxin protein to  cleave the DNA of host cell  to avoid the bacterial escape. We test the function of this toxin protein in E. coli. The result is as follow.


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]


2020 SZPT-CHINA

Result

We streaked the engineered strain into a 1Mm IPTG plate for overnight culture, and observed that the engineered strain containing the RalR gene did not grow on the plate. The expression of RalR was good under the positive and negative controls, and the colony situation in the corresponding area was significantly less than that of the empty vector control group. The result is as follow.

T--SZPT-China--RalR.png

References

Guo Y, Quiroga C, Chen Q, et al. RalR (a DNase) and RalA (a small RNA) form a type I toxin-antitoxin system in Escherichia coli. Nucleic Acids Res. 2014;42(10):6448-6462.