Difference between revisions of "Part:BBa K3771012"

Line 18: Line 18:
  
 
<!-- -->
 
<!-- -->
 +
 +
<br><b style="font-size:1.3rem">Usage and Biology</b>
 +
<br>
 +
<br>The DNA sequence of CS was synthesized by IDT and was amplified by PCR. The Agarose gel electrophoresis result is shown in Fig.2.The part has been confirmed by sequencing and has no mutations.
 +
<br>
 +
 +
<html><div style="width=100%; display:flex; align-items: center; justify-content: center;">
 +
<img src="https://2021.igem.org/wiki/images/f/fb/T--NCKU_Tainan--CS_PCR.png" style="width:35%;">
 +
</div></html>
 +
<p align="center">Fig.2. The electrophoresis result of CS fragment from PCR. M: Marker; Lane 1-3: <i>cs-6xHis</i> PCR product (1312 bp).
 +
</p>
 +
 +
<br>In order to use anti-polyhistidine-tag antibodies to detect the production of CS by western blot, we add 6xHis-tag at the C-terminal of CS. Combining CS and CSAD, we are able to produce taurine from O-phospho-L-serine.
 +
<br>
 +
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3771012 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3771012 SequenceAndFeatures</partinfo>

Revision as of 14:59, 17 October 2021


CS-6xHis


Description


L-cysteine sulfonic acid synthase (CS, L-cysteate synthase) is an enzyme consisting of 419 amino acids, and its size is about 45.85 kDa. It catalyzes the conversion of O-phospho-L-serine and sulfite into L-cysteine sulfonic acid (L-cysteate) and phosphate (Fig.1). Moreover, CS does not catalyze the production of L-cysteine due to its specificity to sulfite[1].

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 and Biology

The DNA sequence of CS was synthesized by IDT and was amplified by PCR. The Agarose gel electrophoresis result is shown in Fig.2.The part has been confirmed by sequencing and has no mutations.

Fig.2. The electrophoresis result of CS fragment from PCR. M: Marker; Lane 1-3: cs-6xHis PCR product (1312 bp).


In order to use anti-polyhistidine-tag antibodies to detect the production of CS by western blot, we add 6xHis-tag at the C-terminal of CS. Combining CS and CSAD, we are able to produce taurine from O-phospho-L-serine.

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 493