Difference between revisions of "Part:BBa K3771079"

 
Line 3: Line 3:
 
<partinfo>BBa_K3771079 short</partinfo>
 
<partinfo>BBa_K3771079 short</partinfo>
  
−
test
+
<br><b style="font-size:1.3rem">Description</b>
 +
<br>
 +
 
 +
<br><i>P<sub>T7</sub>-jju</i> is a composite part consisting of the <i>T7</i> promoter and the <i>cs</i> sequences. This part was used in <i>in vivo</i> testing of taurine production. The sequence for <i>jju</i> and <i>T7</i> promoter were ligated and transformed into <i>E. coli</i> to calculate taurine production using high-performance liquid chromatography (HPLC).
 +
<br>
 +
 
 +
<br><b style="font-size:1.3rem">Biology</b>
 +
<br>
 +
 
 +
<br><i>T7</i> promoter constitutively facilitates the expression of JJU enzyme.
 +
<br>
 +
 
 +
<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:35%;">
 +
</div>
 +
<p align="center">圖片描述</p>
 +
 
 +
<div style="width=100%; display:flex; align-items: center; justify-content: center;">
 +
<img src="https://2021.igem.org/wiki/images/0/09/T--NCKU_Tainan--jju-fragment-pcr.png" style="width:35%;">
 +
</div>
 +
<p align="center">Fig 1. Confirmation of <i>jju</i> fragment by PCR. M: Marker; Lane 1: <i>jju</i> (1180bp)</p>
 +
 
 +
<div style="width=100%; display:flex; align-items: center; justify-content: center;">
 +
<img src="https://2021.igem.org/wiki/images/e/ec/T--NCKU_Tainan--jju-vactor-digestion.png" style="width:35%;">
 +
</div>
 +
<p align="center">Fig 2. Confirmation of pET28a fragment by digestion. M: Marker; Lane 1: pET28a (~5280 bp)</p>
 +
 
 +
<div style="width=100%; display:flex; align-items: center; justify-content: center;">
 +
<img src="https://2021.igem.org/wiki/images/4/44/T--NCKU_Tainan--jju-plate%28DH5a%29.png" style="width:35%;">
 +
</div>
 +
<p align="center">Fig. 3 Transformation/JJU in DH5α</p>
 +
 
 +
<div style="width=100%; display:flex; align-items: center; justify-content: center;">
 +
<img src="https://2021.igem.org/wiki/images/c/c2/T--NCKU_Tainan--jjudigestion.png" style="width:35%;">
 +
</div>
 +
<p align="center">Fig. 4 Confirmation of  pET28a-T7-JJU by digestion.
 +
M: Marker; Lane 1: Colony of pET28a-T7-JJU (~6466 bp)
 +
</p>
 +
 
 +
<div style="width=100%; display:flex; align-items: center; justify-content: center;">
 +
<img src="https://2021.igem.org/wiki/images/e/e6/T--NCKU_Tainan--jju-plate%28BL21%28DE3%29%29.png" style="width:35%;">
 +
</div>
 +
<p align="center">Fig. 5 Transform into BL21(DE3)</p>
 +
 
 +
<div style="width=100%; display:flex; align-items: center; justify-content: center;">
 +
<img src="圖片網址" style="width:35%;">
 +
</div>
 +
<p align="center">Fig. 6 Confirmation of protein expression of JJU.
 +
M: Marker; Lane1: whole cell of JJU in BL21(DE3); Lane2: soluble protein of JJUBL21(DE3) (~43 kDa)
 +
</p>
 +
 
 +
<div style="width=100%; display:flex; align-items: center; justify-content: center;">
 +
<img src="https://2021.igem.org/wiki/images/1/1b/T--NCKU_Tainan--jju-plate%28BD7G%29.png" style="width:35%;">
 +
</div>
 +
<p align="center">Fig. 7 Transform into BD7G</p>
 +
 
 +
<div style="width=100%; display:flex; align-items: center; justify-content: center;">
 +
<img src="圖片網址" style="width:35%;">
 +
</div>
 +
<p align="center">Fig. 8 Confirmation of protein expression of JJU.
 +
M: Marker; Lane1: whole cell of JJU in BD7G; Lane2: soluble protein of JJU in BD7G(~43 kDa)
 +
</p>
 +
 
 +
<br><b style="font-size:1.3rem">Usage</b>
 +
<br>
 +
 
 +
<br><b style="font-size:1.3rem">Characterization</b>
 +
<br>
 +
 
 +
<br><b style="font-size:1.3rem">References</b>
 +
<br>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 21:35, 17 October 2021


T7-JJU


Description


PT7-jju is a composite part consisting of the T7 promoter and the cs sequences. This part was used in in vivo testing of taurine production. The sequence for jju and T7 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 JJU enzyme.

<img src="T--NCKU_Tainan--taurine_pathway_1.png" style="width:35%;">

圖片描述

<img src="T--NCKU_Tainan--jju-fragment-pcr.png" style="width:35%;">

Fig 1. Confirmation of jju fragment by PCR. M: Marker; Lane 1: jju (1180bp)

<img src="T--NCKU_Tainan--jju-vactor-digestion.png" style="width:35%;">

Fig 2. Confirmation of pET28a fragment by digestion. M: Marker; Lane 1: pET28a (~5280 bp)

<img src="T--NCKU_Tainan--jju-plate%28DH5a%29.png" style="width:35%;">

Fig. 3 Transformation/JJU in DH5α

<img src="T--NCKU_Tainan--jjudigestion.png" style="width:35%;">

Fig. 4 Confirmation of pET28a-T7-JJU by digestion. M: Marker; Lane 1: Colony of pET28a-T7-JJU (~6466 bp)

<img src="T--NCKU_Tainan--jju-plate%28BL21%28DE3%29%29.png" style="width:35%;">

Fig. 5 Transform into BL21(DE3)

<img src="圖片網址" style="width:35%;">

Fig. 6 Confirmation of protein expression of JJU. M: Marker; Lane1: whole cell of JJU in BL21(DE3); Lane2: soluble protein of JJUBL21(DE3) (~43 kDa)

<img src="T--NCKU_Tainan--jju-plate%28BD7G%29.png" style="width:35%;">

Fig. 7 Transform into BD7G

<img src="圖片網址" style="width:35%;">

Fig. 8 Confirmation of protein expression of JJU. M: Marker; Lane1: whole cell of JJU in BD7G; Lane2: soluble protein of JJU in BD7G(~43 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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]