Difference between revisions of "Part:BBa K3771077"

 
 
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<partinfo>BBa_K3771077 short</partinfo>
 
<partinfo>BBa_K3771077 short</partinfo>
  
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<br><b style="font-size:1.3rem">Description</b>
  
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<br><i>P<sub>trc</sub>-csad</i> is a composite part consisting of the <i>trc</i> promoter and the <i>csad</i> sequences. This part was used in <i>in vivo</i> testing of taurine production. The sequence for <i>csad</i> and <i>trc</i> promoter were ligated and transformed into <i>E. coli</i> to calculate taurine production using high-performance liquid chromatography (HPLC).
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<br><b style="font-size:1.3rem">Biology</b>
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<br><i>trc</i> promoter constitutively facilitates the expression of CSAD enzyme.
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<br>
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<div style="width=100%; display:flex; align-items: center; justify-content: center;">
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<img src="https://2021.igem.org/wiki/images/c/c9/T--NCKU_Tainan--taurine_pathway_1.png" style="width:60%;">
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  <br>
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  <p align="center">Fig. 1. Taurine pathways in <i>E. coli</i>
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  </p>
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<br><b style="font-size:1.3rem">Usage</b>
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  <br>CSAD was used in <i>in vivo</i> testing of taurine production. The sequence for CDO1 enzyme and <i>trc</i> promoter were ligated and transformed into <i>E. coli</i> to calculate taurine production using high-performance liquid chromatography (HPLC). <br>
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<br><b style="font-size:1.3rem">Characterization</b>
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<br>
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<div style="width=100%; display:flex; align-items: center; justify-content: center;">
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<img src="https://2021.igem.org/wiki/images/9/92/T--NCKU_Tainan--CSAD2-PCR.png" style="width:35%;">
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</div>
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  <p align="center">Fig. 2. Confirmation of <i>csad</i> fragment by PCR. M: Marker; Lane 1: <i>csad</i> (1368 bp)</p>
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<div style="width=100%; display:flex; align-items: center; justify-content: center;">
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<img src="https://2021.igem.org/wiki/images/2/26/T--NCKU_Tainan--CSAD2-digestion.png" style="width:35%;">
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</div>
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  <p align="center">Fig. 3. Confirmation of  pSUI-<i>P<sub>trc</sub>-csad</i> by digestion.
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    M: Marker; Lane 1~3: Different colonies of pSUI-<i>P<sub>trc</sub>-csad</i> (3674 bp)
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</p>
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<div style="width=100%; display:flex; align-items: center; justify-content: center;">
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<img src="https://2021.igem.org/wiki/images/c/cd/T--NCKU_Tainan--CSAD2-plate%28DH5a%29.png" style="width:35%;">
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</div>
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<p align="center">Fig. 4. Transformation/CSAD in DH5α</p>
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<div style="width=100%; display:flex; align-items: center; justify-content: center;">
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<img src="https://2021.igem.org/wiki/images/9/92/T--NCKU_Tainan--CSAD-PAGE%28DH5a%29.png" style="width:35%;">
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</div>
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<p align="center">Fig. 5. Confirmation of protein expression of CSAD.
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M: Marker; Lane 1: whole cell of CSAD in DH5α; Lane 2: soluble protein of CSAD in DH5α (~50 kDa)
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</p>
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<br><b style="font-size:1.3rem">References</b>
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Latest revision as of 03:16, 22 October 2021


Ptrc-CSAD


Description
Ptrc-csad is a composite part consisting of the trc promoter and the csad sequences. This part was used in in vivo testing of taurine production. The sequence for csad and trc promoter were ligated and transformed into E. coli to calculate taurine production using high-performance liquid chromatography (HPLC).

Biology
trc promoter constitutively facilitates the expression of CSAD enzyme.


Fig. 1. Taurine pathways in E. coli


Usage
CSAD was used in in vivo testing of taurine production. The sequence for CDO1 enzyme and trc promoter were ligated and transformed into E. coli to calculate taurine production using high-performance liquid chromatography (HPLC).

Characterization

Fig. 2. Confirmation of csad fragment by PCR. M: Marker; Lane 1: csad (1368 bp)

Fig. 3. Confirmation of pSUI-Ptrc-csad by digestion. M: Marker; Lane 1~3: Different colonies of pSUI-Ptrc-csad (3674 bp)

Fig. 4. Transformation/CSAD in DH5α

Fig. 5. Confirmation of protein expression of CSAD. M: Marker; Lane 1: whole cell of CSAD in DH5α; Lane 2: soluble protein of CSAD in DH5α (~50 kDa)


References
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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 337
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 122