Difference between revisions of "Part:BBa K3771008"
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<br>CoaBC is an enzyme that weighs 55.4 kDa. CoaBC functions in the JJU10-CoaBC taurine biosynthesis pathway, converting L-cystate to taurine. [3]<br>
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<br>CoaBC is an enzyme that weighs 55.4 kDa. CoaBC functions in the JJU10-CoaBC taurine biosynthesis pathway, converting L-cystate to taurine. [3]
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<br>CoaBC enzyme was used in vitro testing of taurine production. The sequence for CoaBC enzyme and trc promoter were ligated and transformed into E. coli to calculate taurine production using high-performance liquid chromatography (HPLC).  
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<br>CoaBC was used in vitro testing of taurine production. The sequence for CoaBC 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.  
 
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<br><b style="font-size:1.3rem">Characterization
 
<br><b style="font-size:1.3rem">Characterization

Revision as of 07:16, 17 October 2021


CSAD-6xHis


Description

CoaBC is an enzyme that weighs 55.4 kDa. CoaBC functions in the JJU10-CoaBC taurine biosynthesis pathway, converting L-cystate to taurine. [3]


Biology


<img src="Biology image" style="width:35%;">



CoaBC is an enzyme in the JJU10-CoaBC pathway, one of three possible taurine synthesis pathways. Its main function is to convert L-cystate to taurine.


Usage

CoaBC was used in vitro testing of taurine production. The sequence for CoaBC 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.


Characterization

The CoaBC fragment was synthesized by IDT and amplified by PCR. Agarose gel electrophoresis result is shown in Fig. 2.


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 250
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 35