Difference between revisions of "Part:BBa K3771080"

 
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<partinfo>BBa_K3771080 short</partinfo>
 
<partinfo>BBa_K3771080 short</partinfo>
  
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<br><b style="font-size:1.3rem">Description</b>
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<br><i>P<sub>T7</sub>-coaBC</i> is a composite part consisting of the <i>T7</i> promoter and the <i>coaBC</i> sequences. This part was used in <i>in vivo</i> testing of taurine production. The sequence for <i>jju</i> and <i>coaBC</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>
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<br>
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<br><i>T7</i> promoter constitutively facilitates the expression of CoaBC enzyme.
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<br>
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<div style="width=100%; display:flex; align-items: center; justify-content: center;">
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<img src="https://2021.igem.org/wiki/images/c/c9/T--NCKU_Tainan--taurine_pathway_1.png" style="width:35%;">
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</div>
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<p align="center">圖片描述</p>
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<div style="width=100%; display:flex; align-items: center; justify-content: center;">
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<img src="https://2021.igem.org/wiki/images/7/7f/T--NCKU_Tainan--CoaBC-PCR.png" style="width:35%;">
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</div>
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<p align="center">Fig. 1 Confirmation of coaBC fragment by PCR. M: Marker; Lane 1: coaBC (1221 bp)</p>
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<div style="width=100%; display:flex; align-items: center; justify-content: center;">
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<img src="https://2021.igem.org/wiki/images/5/53/T--NCKU_Tainan--CoaBC-plate%28DH5a%29.png" style="width:35%;">
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</div>
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<p align="center">Fig. 2 Transformation/CoaBC in DH5α</p>
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<div style="width=100%; display:flex; align-items: center; justify-content: center;">
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<img src="https://2021.igem.org/wiki/images/5/59/T--NCKU_Tainan--CoaBC-digestion.png" style="width:35%;">
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</div>
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<p align="center">Fig. 3 Confirmation of  pET28a-T7-CoaBC by digestion.
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M: Marker; Lane 1~3: Different colonies of pET28a-T7-CoaBC (5398 bp)
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</p>
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<div style="width=100%; display:flex; align-items: center; justify-content: center;">
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<img src="https://2021.igem.org/wiki/images/d/d1/T--NCKU_Tainan--CoaBC-plate%28BL21%28DE3%29%29.png" style="width:35%;">
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</div>
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<p align="center">Fig. 4 Transform into BL21(DE3)</p>
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<div style="width=100%; display:flex; align-items: center; justify-content: center;">
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<img src="https://2021.igem.org/wiki/images/3/39/T--NCKU_Tainan--CoaBC-PAGE.png" style="width:35%;">
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</div>
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<p align="center">Fig. 5 Confirmation of protein expression of CoaBC.
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M: Marker; Lane1: whole cell of CoaBC in BL21(DE3); Lane2: soluble protein of CoaBC in BL21(DE3) (~45 kDa)
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</p>
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<br><b style="font-size:1.3rem">Usage</b>
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<br>
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<br><b style="font-size:1.3rem">Characterization</b>
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<br>
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<br><b style="font-size:1.3rem">References</b>
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<br>
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Revision as of 21:44, 17 October 2021


T7-CoaBC


Description

PT7-coaBC is a composite part consisting of the T7 promoter and the coaBC sequences. This part was used in in vivo testing of taurine production. The sequence for jju and coaBC promoter were ligated and transformed into E. coli to calculate taurine production using high-performance liquid chromatography (HPLC).

Biology

T7 promoter constitutively facilitates the expression of CoaBC enzyme.

圖片描述

Fig. 1 Confirmation of coaBC fragment by PCR. M: Marker; Lane 1: coaBC (1221 bp)

Fig. 2 Transformation/CoaBC in DH5α

Fig. 3 Confirmation of pET28a-T7-CoaBC by digestion. M: Marker; Lane 1~3: Different colonies of pET28a-T7-CoaBC (5398 bp)

Fig. 4 Transform into BL21(DE3)

Fig. 5 Confirmation of protein expression of CoaBC. M: Marker; Lane1: whole cell of CoaBC in BL21(DE3); Lane2: soluble protein of CoaBC in BL21(DE3) (~45 kDa)


Usage

Characterization

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 NgoMIV site found at 632
  • 1000
    COMPATIBLE WITH RFC[1000]