Difference between revisions of "Part:BBa K3645011"
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===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K3645011 parameters</partinfo>
 
<partinfo>BBa_K3645011 parameters</partinfo>
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==Contribution From XMU-China 2020==
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'''Group''': [https://2020.igem.org/Team/XMU-China iGEM Team XMU-China 2020]
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'''Author''': Shi Zhang
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'''Summary''': colony forming unit (CFU)
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===Characterization from iGEM20-XMU-China===
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;MazF is toxin protein in MazF-MazE, toxin-antitoxin module. It was first registered in 2013 and used as an mRNA endonuclease. It kills bacteria without cracking them, which will directly influence the value of OD<sub>600</sub>.
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;In order to quantify the toxicity of mazF, colony forming unit (CFU) cell viability assays were used to measure functionality of the circuit(1).
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Because direct expression of MazF will kill bacteria, we construct the circuit "P<sub>Bad/araC</sub>-RBS-MazF-terminator-pSB1C3"(<partinfo>BBa_K3332083</partinfo>) and "P<sub>Bad/araC</sub>-RBS-EYFP-terminator-pSB1C3"(<partinfo>BBa_K3332082</partinfo>) in ''E.coli'' BL21(DE3) to characterize its function.
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<html>
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    <figure>
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        <img src="https://2020.igem.org/wiki/images/0/0e/T--XMU-China--XMU-China_2020_pBAD_B0034_mazF_B0015.png" width="40%" style="float:center">
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        <figcaption>
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        <p style="font-size:1rem">
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        </p>
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        </figcaption>
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    </figure>
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</html>
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:'''Fig.1''' genetic circuit of P<sub>Bad/araC</sub>-RBS-MazF-terminator.
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;When 0.2% arabinose was added, the experimental group showed a sharp decrease on the CFU, which indicates the protein encoded by MazF is fatal to ''E. coli''. It further demonstrated that the MazF encoded by J23100-RBS (<partinfo>BBa_K880005</partinfo>) was able to work as a toxin.
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<html>
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    <figure>
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        <img src="https://2020.igem.org/wiki/images/c/c7/T--XMU-China--XMU-China_2020_BM-CFU.png" width="45%" style="float:center">
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        <figcaption>
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        <p style="font-size:1rem">
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        </p>
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        </figcaption>
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    </figure>
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</html>
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:'''Fig.2''' The results of CFU.The round dot indicates the non induction group, and the square dot indicates the induction group.
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Revision as of 13:15, 17 October 2021


Target-AID (CBE)

Contains the full CDS of Target-AID, whose Cas9 part was replace with our lab's dCas9.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1099
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 4775
    Illegal BamHI site found at 3378
    Illegal XhoI site found at 4384
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Contribution From XMU-China 2020

Group: iGEM Team XMU-China 2020

Author: Shi Zhang

Summary: colony forming unit (CFU)

Characterization from iGEM20-XMU-China

        MazF is toxin protein in MazF-MazE, toxin-antitoxin module. It was first registered in 2013 and used as an mRNA endonuclease. It kills bacteria without cracking them, which will directly influence the value of OD600.

        In order to quantify the toxicity of mazF, colony forming unit (CFU) cell viability assays were used to measure functionality of the circuit(1).

        Because direct expression of MazF will kill bacteria, we construct the circuit "PBad/araC-RBS-MazF-terminator-pSB1C3"(BBa_K3332083) and "PBad/araC-RBS-EYFP-terminator-pSB1C3"(BBa_K3332082) in E.coli BL21(DE3) to characterize its function.

Fig.1 genetic circuit of PBad/araC-RBS-MazF-terminator.

        When 0.2% arabinose was added, the experimental group showed a sharp decrease on the CFU, which indicates the protein encoded by MazF is fatal to E. coli. It further demonstrated that the MazF encoded by J23100-RBS (BBa_K880005) was able to work as a toxin.

Fig.2 The results of CFU.The round dot indicates the non induction group, and the square dot indicates the induction group.