Difference between revisions of "Part:BBa J09855:Experience"

(User Reviews)
Line 30: Line 30:
 
Also, we add RFP at the downstream of Plux promoter. This fluorescence protein is very easy to detect so that we can use this part to test AHL and work as reporter. Our new part is BBa_K766003.
 
Also, we add RFP at the downstream of Plux promoter. This fluorescence protein is very easy to detect so that we can use this part to test AHL and work as reporter. Our new part is BBa_K766003.
  
This part is tested by '''ETH Zürich-iGEM 2013'''<br>
+
<br><br>This part is tested by '''ETH Zürich-iGEM 2013'''<br>
Leaky expression of plac promoter
+
The BBa_J09855 was cloned together with GFP [https://parts.igem.org/Part:BBa_E0840 BBa_E0840] to build a sensor construct for AHL. The construct was tested in LB liquid cultures over 16 h without and with induction through AHL (100 nM). The GFP fluorescence was tested using a TECAN plate reader and normalized to the OD<sub>600</sub>.
 +
 
 +
[[File:ETHZ_J09855registryentry.png|center|thumb| <b>The normalized GFP fluorescence of the GFP receiver construct based on BBa_J09855.</b>]]

Revision as of 12:26, 30 October 2013

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_J09855

This part is tested by 2012 iGEM team Tsinghua.

We use this part to work as a promoter system which can be activated by C6HSL. We can add some other gene dowmstream so that AHL can work as a signal. In our project this year, we use this part and modified it to get more complicated construct.

User Reviews

UNIQ20b89436c0510619-partinfo-00000000-QINU UNIQ20b89436c0510619-partinfo-00000001-QINU This part is tested by 2012 iGEM team Tsinghua.

To tell the truth, when we use this part for the first time, it didn't work well. Then we sequenced the part using prefix and suffix primer. We found that there are some part missed in Plac promoter at the beginning and in Plux promoter at the end. Maybe this is why it can't work.


We improve this part and modify sequence. We add several nucleic acid to the Plac promoter and plux promoter. This modification improve the efficiency of transcription. The sequensing results show that our construct is correct.

Also, we add RFP at the downstream of Plux promoter. This fluorescence protein is very easy to detect so that we can use this part to test AHL and work as reporter. Our new part is BBa_K766003.



This part is tested by ETH Zürich-iGEM 2013
The BBa_J09855 was cloned together with GFP BBa_E0840 to build a sensor construct for AHL. The construct was tested in LB liquid cultures over 16 h without and with induction through AHL (100 nM). The GFP fluorescence was tested using a TECAN plate reader and normalized to the OD600.

The normalized GFP fluorescence of the GFP receiver construct based on BBa_J09855.