Difference between revisions of "Part:BBa J09855:Experience"
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The BBa_J09855 was used as a LuxR-generating construct together with GFP [https://parts.igem.org/Part:BBa_E0840 BBa_E0840] expressed from P<sub>LuxR</sub> [https://parts.igem.org/Part:BBa_R0062 BBa_R0062] to build a sensor system 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 measured using a TECAN plate reader and normalized to the OD<sub>600</sub>. | The BBa_J09855 was used as a LuxR-generating construct together with GFP [https://parts.igem.org/Part:BBa_E0840 BBa_E0840] expressed from P<sub>LuxR</sub> [https://parts.igem.org/Part:BBa_R0062 BBa_R0062] to build a sensor system 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 measured using a TECAN plate reader and normalized to the OD<sub>600</sub>. | ||
| − | [[File:ETHZ J09855registryentry2.png| | + | [[File:ETHZ J09855registryentry2.png|900px|center|thumb| <b>The normalized GFP fluorescence of the AHL inducible GFP system based on BBa_J09855.</b>]] |
Revision as of 13:54, 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
UNIQ5cbd1d7950851ab8-partinfo-00000000-QINU UNIQ5cbd1d7950851ab8-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 was tested by ETH Zurich-iGEM 2013
The BBa_J09855 was used as a LuxR-generating construct together with GFP BBa_E0840 expressed from PLuxR BBa_R0062 to build a sensor system 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 measured using a TECAN plate reader and normalized to the OD600.
