Difference between revisions of "Part:BBa K3771077"
Line 15: Line 15:
 
<br><i>trc</i> promoter constitutively facilitates the expression of CSAD enzyme.
 
<br><i>trc</i> promoter constitutively facilitates the expression of CSAD enzyme.
 
<br>
 
<br>
−
 
 
<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:50%;">
+
<img src="https://2021.igem.org/wiki/images/c/c9/T--NCKU_Tainan--taurine_pathway_1.png" style="width:35%;">
 
</div>
 
</div>
 +
  <br>
 +
  <p align="center">Fig. 1 L-cysteine sulfonic acid synthase catalyzes the reaction that turns O-phospho-L-serine into L-cysteine sulfonic acid. CDO1, L-cysteine dioxygenase; CSAD, L-cysteine sulfinic acid decarboxylase
  
  
Line 42: Line 43:
 
<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. 1 Confirmation of csad fragment by PCR. M: Marker; Lane 1: csad (1368 bp)</p>
+
  <p align="center">Fig. 2 Confirmation of csad 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. 2 Confirmation of  pSUI-Ptrc-CSAD by digestion.
+
  <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-Ptrc-CSAD (3674 bp)
+
    M: Marker; Lane 1~3: Different colonies of pSUI-<i>P<sub>trc</sub>-csad</i> (3674 bp)
 
</p>
 
</p>
  

Revision as of 01:41, 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 L-cysteine sulfonic acid synthase catalyzes the reaction that turns O-phospho-L-serine into L-cysteine sulfonic acid. CDO1, L-cysteine dioxygenase; CSAD, L-cysteine sulfinic acid decarboxylase
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. 3 Transformation/CSAD in DH5α

Fig. 4 Confirmation of protein expression of CSAD. M: Marker; Lane1: whole cell of CSAD in DH5α; Lane2: soluble protein of CSAD in DH5α (~22 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