Difference between revisions of "Part:BBa K3771077"
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<br><b style="font-size:1.3rem">Description</b> | <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). | <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> | <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. | <br><i>trc</i> promoter constitutively facilitates the expression of CSAD enzyme. | ||
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<div style="width=100%; display:flex; align-items: center; justify-content: center;"> | <div style="width=100%; display:flex; align-items: center; justify-content: center;"> | ||
| − | <img src="https://2021.igem.org/wiki/images/c/c9/T--NCKU_Tainan--taurine_pathway_1.png" style="width: | + | <img src="https://2021.igem.org/wiki/images/c/c9/T--NCKU_Tainan--taurine_pathway_1.png" style="width:60%;"> |
</div> | </div> | ||
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| + | <p align="center">Fig. 1. Taurine pathways in <i>E. coli</i> | ||
| + | </p> | ||
<br><b style="font-size:1.3rem">Usage</b> | <br><b style="font-size:1.3rem">Usage</b> | ||
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| − | <br>CSAD was used in in vivo 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> | + | <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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<img src="https://2021.igem.org/wiki/images/9/92/T--NCKU_Tainan--CSAD2-PCR.png" style="width:35%;"> | <img src="https://2021.igem.org/wiki/images/9/92/T--NCKU_Tainan--CSAD2-PCR.png" style="width:35%;"> | ||
</div> | </div> | ||
| − | <p align="center">Fig. | + | <p align="center">Fig. 2. Confirmation of <i>csad</i> fragment by PCR. M: Marker; Lane 1: <i>csad</i> (1368 bp)</p> |
<div style="width=100%; display:flex; align-items: center; justify-content: center;"> | <div style="width=100%; display:flex; align-items: center; justify-content: center;"> | ||
<img src="https://2021.igem.org/wiki/images/2/26/T--NCKU_Tainan--CSAD2-digestion.png" style="width:35%;"> | <img src="https://2021.igem.org/wiki/images/2/26/T--NCKU_Tainan--CSAD2-digestion.png" style="width:35%;"> | ||
</div> | </div> | ||
| − | <p align="center">Fig. | + | <p align="center">Fig. 3. Confirmation of pSUI-<i>P<sub>trc</sub>-csad</i> by digestion. |
| − | M: Marker; Lane 1~3: Different colonies of pSUI- | + | M: Marker; Lane 1~3: Different colonies of pSUI-<i>P<sub>trc</sub>-csad</i> (3674 bp) |
</p> | </p> | ||
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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%;"> | <img src="https://2021.igem.org/wiki/images/c/cd/T--NCKU_Tainan--CSAD2-plate%28DH5a%29.png" style="width:35%;"> | ||
</div> | </div> | ||
| − | <p align="center">Fig. | + | <p align="center">Fig. 4. Transformation/CSAD in DH5α</p> |
<div style="width=100%; display:flex; align-items: center; justify-content: center;"> | <div style="width=100%; display:flex; align-items: center; justify-content: center;"> | ||
<img src="https://2021.igem.org/wiki/images/9/92/T--NCKU_Tainan--CSAD-PAGE%28DH5a%29.png" style="width:35%;"> | <img src="https://2021.igem.org/wiki/images/9/92/T--NCKU_Tainan--CSAD-PAGE%28DH5a%29.png" style="width:35%;"> | ||
</div> | </div> | ||
| − | <p align="center">Fig. | + | <p align="center">Fig. 5. Confirmation of protein expression of CSAD. |
| − | M: Marker; | + | M: Marker; Lane 1: whole cell of CSAD in DH5α; Lane 2: soluble protein of CSAD in DH5α (~50 kDa) |
</p> | </p> | ||
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:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 337
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 122