Difference between revisions of "Part:BBa K3771041"
 
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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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  <p align="center">Figure.1 Taurine pathways in <i>E. coli</i> [1,2].</p>
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  <p align="center">Fig. 1. Taurine pathways in <i>E. coli</i> [1,2].</p>
  
 
    
 
    
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<br><b style="font-size:1.3rem">Usage
 
<br><b style="font-size:1.3rem">Usage
 
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<br>CDO1 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). A 6xHis-tag is added to the C-terminal of the CoaBC protein, allowing for confirmation of JJU10 expression by western blot using the anti-6xHis antibody.<br>
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<br>CDO1 was used in <i>in vivo</i> testing of taurine production. The sequence for CDO1 enzyme and trc 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">Characterization  
 
<br><b style="font-size:1.3rem">Characterization  
 
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<br>The CDO1 fragment was synthesized by IDT and amplified by PCR. Agarose gel electrophoresis result is shown in Fig. 2.<br>  
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<br>The CDO1 fragment was synthesized by IDT and amplified by PCR. Agarose gel electrophoresis result is shown in Fig. 2.<br>
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  <p align="center">Fig. 2. PCR confirmation of <i>cdo1-6xHis</i></p> 
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<br>CDO1 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). A 6xHis-tag is added to the C-terminal of the CDO1 protein, allowing for confirmation of CDO1 expression by western blot using the anti-6xHis antibody.<br>
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<br><b style="font-size:1.3rem">References
 
<br><b style="font-size:1.3rem">References
 
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<br>Joo Y-C, Ko YJ, You SK, et al. Creating a New Pathway in Corynebacterium glutamicum for the Production of Taurine as a Food Additive. Journal of Agricultural and Food Chemistry. 2018;66(51):13454-13463. doi:10.1021/acs.jafc.8b05093<br>
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<br>1. Joo Y-C, Ko YJ, You SK, et al. Creating a New Pathway in Corynebacterium glutamicum for the Production of Taurine as a Food Additive. Journal of Agricultural and Food Chemistry. 2018;66(51):13454-13463. doi:10.1021/acs.jafc.8b05093<br>
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===Usage and Biology===
 
===Usage and Biology===
  

Latest revision as of 03:35, 22 October 2021


CDO1-6xHis


Description

L-cysteine dioxygenase (CDO1) is an enzyme that weighs 23 kDa. CDO1 functions in the L-cysteine sulfinic acid taurine biosynthesis pathway, converting L-cysteine to L-cysteine sulfinic acid. [1]

Fig. 1. Taurine pathways in E. coli [1,2].



CDO1 is part of the L-cysteine sulfinic acid pathway, one of three possible taurine synthesis pathways. CDO1 oxidizes L-cysteine naturally found in the intestine into L-cysteine sulfinic acid, which is later converted to taurine by CSAD [1].



Usage


CDO1 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. PCR confirmation of cdo1-6xHis




CDO1 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). A 6xHis-tag is added to the C-terminal of the CDO1 protein, allowing for confirmation of CDO1 expression by western blot using the anti-6xHis antibody.




References
1. Joo Y-C, Ko YJ, You SK, et al. Creating a New Pathway in Corynebacterium glutamicum for the Production of Taurine as a Food Additive. Journal of Agricultural and Food Chemistry. 2018;66(51):13454-13463. doi:10.1021/acs.jafc.8b05093

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 NgoMIV site found at 447
  • 1000
    COMPATIBLE WITH RFC[1000]