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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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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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.
  
 
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==2020 SZPT-CHINA==
 
==2020 SZPT-CHINA==
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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===
 
===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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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 shown in the following.
 
[[File:T--SZPT-China--RalR.png|thumb|center|300px|pET28a(+):vector positive control;ddH2O:negative control;RalR:Non-specific endonuclease]]
 
[[File:T--SZPT-China--RalR.png|thumb|center|300px|pET28a(+):vector positive control;ddH2O:negative control;RalR:Non-specific endonuclease]]
  
 
===References===
 
===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.
 
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.

Revision as of 08:49, 20 October 2020


Non-specific endonuclease

Description

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.

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 shown in the following.

pET28a(+):vector positive control;ddH2O:negative control;RalR:Non-specific endonuclease

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.