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