Difference between revisions of "Part:BBa K3722007"

 
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A part for TnaA expression in the preliminary experiment.
 
A part for TnaA expression in the preliminary experiment.
  
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===Usage and Biology===
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===Biology===
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TnaA cleaves 6-Br-Trp into 6-Br-indole, pyruvate and ammonia, so we can use this composite part to synthetic 6-Br-indole as the second step to product 6,6’-dibromo indigo.
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<html>
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    <figure>
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        <img src="https://2021.igem.org/wiki/images/e/e9/T--NWU-CHINA-A--part001.png" width="20%" 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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===Usage===
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We construct the composite part <partinfo>BBa_K3332052</partinfo> and transformed it into E.coli DH5α& E. coli BL21 (DE3) to verify its expression.
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===Characterization===
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1.Identification
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After plasmid construction, we performed PCR to certify and the resulting was shown down.
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<html>
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    <figure>
 +
        <img src="https://2021.igem.org/wiki/images/2/2d/T--NWU-CHINA-A--part071.png" width="30%" 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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2.The proof of expression
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We used IPTG to induce the expression at different times.
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<html>
 +
    <figure>
 +
        <img src="https://2021.igem.org/wiki/images/f/f7/T--NWU-CHINA-A--part072.png" width="60%" style="float:center">
 +
        <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'''SDS-PAGE analysis of TnaA .Induction condition: LB media, OD600 0.6 ~ 0.8, IPTG 0.1 mM, 37 °C and 200 rpm.(1:negitive control  2: 9h  3:10.5h  4:12h  5:13.5h  6:15h  7:16.5h    8: 18h    9: 19.5h)
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===References===
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Kim, H. J. et al. Biosynthesis of indigo in Escherichia coli expressing self- sufficient CYP102A from Streptomyces cattleya. Dyes Pigments 140, 29–35 (2017).
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Latest revision as of 00:12, 22 October 2021


T7_Promoter-RBS-TnaA-T7_Terminator

A part for TnaA expression in the preliminary experiment.


Biology

TnaA cleaves 6-Br-Trp into 6-Br-indole, pyruvate and ammonia, so we can use this composite part to synthetic 6-Br-indole as the second step to product 6,6’-dibromo indigo.

Usage

We construct the composite part BBa_K3332052 and transformed it into E.coli DH5α& E. coli BL21 (DE3) to verify its expression.


Characterization

1.Identification After plasmid construction, we performed PCR to certify and the resulting was shown down.

2.The proof of expression We used IPTG to induce the expression at different times.

Fig.1SDS-PAGE analysis of TnaA .Induction condition: LB media, OD600 0.6 ~ 0.8, IPTG 0.1 mM, 37 °C and 200 rpm.(1:negitive control 2: 9h 3:10.5h 4:12h 5:13.5h 6:15h 7:16.5h 8: 18h 9: 19.5h)


References

Kim, H. J. et al. Biosynthesis of indigo in Escherichia coli expressing self- sufficient CYP102A from Streptomyces cattleya. Dyes Pigments 140, 29–35 (2017).


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]