Difference between revisions of "Part:BBa J23117:Experience"
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Additionally to our characterization of this part, we also used it for our reporter system and it worked very good! We also improved it by switching the pre- and suffix, basically inverting it. This way, we were able to use it in an "inverted" expression unit on the same vector as our reporter system. For further information see: https://parts.igem.org/wiki/index.php?title=Part:BBa_K1045011 | Additionally to our characterization of this part, we also used it for our reporter system and it worked very good! We also improved it by switching the pre- and suffix, basically inverting it. This way, we were able to use it in an "inverted" expression unit on the same vector as our reporter system. For further information see: https://parts.igem.org/wiki/index.php?title=Part:BBa_K1045011 | ||
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| + | <partinfo>BBa_J23117 StartReviews</partinfo> | ||
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| + | <partinfo>BBa_J23117 AddReview 5</partinfo> | ||
| + | <I>iGEM15_UNITN-Trento</I> | ||
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| + | We used the part BBa_J23117 in the InterLab Measurement context. We assembled a measurement device by inserting the part <html><a href="https://parts.igem.org/Part:BBa_I13504">BBa_I13504</a></html> amplified by PCR into the part BBa_J23117. The device was characterized in <i>E. coli</i> NEB10β, JM109, and NEB Express by measuring GFP expression with a cuvette-based spectrofluorometer, a fluorescence plate reader, and a flow cytometer. We also confirmed the promoter activity with a cell-free S30 extract system and measured mRNA by RT-qPCR. <br> <br> | ||
| + | For a better understanding on protocols and characterizations, please check out our Wiki page <html><a href="http://2015.igem.org/Team:UNITN-Trento/Measurement">UNITN-Trento iGEM 2015</a>! <br> <br></html> | ||
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====Evaluation of Anderson promoter J23117 in ''E. coli'' by [http://2013.igem.org/Team:Goettingen iGEM Göttingen 2013]==== | ====Evaluation of Anderson promoter J23117 in ''E. coli'' by [http://2013.igem.org/Team:Goettingen iGEM Göttingen 2013]==== | ||
Revision as of 16:11, 12 September 2015
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_J23117
Evaluation of Anderson promoter J23117 in B. subtilis by iGEM-Team LMU-Munich 2012
This Anderson promoter was evaluated without fused RFP with the lux operon as a reporter in B. subtilis. See the new BioBrick BBa_K823013 without RFP and have a look at the [http://2012.igem.org/Team:LMU-Munich/Data/Anderson Data] from the evaluation in B. subtilis.
User Reviews
UNIQ53fe99cd3c6b4779-partinfo-00000000-QINU UNIQ53fe99cd3c6b4779-partinfo-00000001-QINU
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iGEM Team Göttingen 2013 |
Additionally to our characterization of this part, we also used it for our reporter system and it worked very good! We also improved it by switching the pre- and suffix, basically inverting it. This way, we were able to use it in an "inverted" expression unit on the same vector as our reporter system. For further information see: https://parts.igem.org/wiki/index.php?title=Part:BBa_K1045011 |
UNIQ53fe99cd3c6b4779-partinfo-00000003-QINU
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iGEM15_UNITN-Trento |
We used the part BBa_J23117 in the InterLab Measurement context. We assembled a measurement device by inserting the part BBa_I13504 amplified by PCR into the part BBa_J23117. The device was characterized in E. coli NEB10β, JM109, and NEB Express by measuring GFP expression with a cuvette-based spectrofluorometer, a fluorescence plate reader, and a flow cytometer. We also confirmed the promoter activity with a cell-free S30 extract system and measured mRNA by RT-qPCR. |
Evaluation of Anderson promoter J23117 in E. coli by [http://2013.igem.org/Team:Goettingen iGEM Göttingen 2013]
Shown here:
Upper two pictures: Growth curves of promoter strains on the left, growth curves of control strains on the right. Three biological replicates are shown.
Middle two pictures: RFP/OD600 of promoter strains on the left, RFP/OD600 of control strains on the right. Three biological replicates are shown.
Bottom three pictures: qRT PCR promoter analyses in three different growth phases. Promoters are normalised against BBa_J23117 .
Promotor 1:BBa_J23117
Promoter 2:BBa_J23116
Promoter 3:BBa_J23110
Promoter 4:BBa_J23118
The promoter strength was measured by using the reporter gene rfp.
Three different approaches were used: 1. RFP measurement, 2. qRT-PCR analyses and 3. single cell microscopy. Moreover, the first and the second approach characterised the promoter activity along the growth curve and to three important time points, respectively.
Taken together, the majority of our results from these approaches showed that BBa_J23117 has the lowest promoter activity compared to BBa_J23116, BBa_J23110 and BBa_J23118. For more details, visit the discussion on our wikipage.
on the left side: bright field (BR), on the right side: RFP filter (rfp)
P1: BBa_J23117, P2: BBa_J23116, P3: BBa_J23110, P4: BBa_J23118, P8: control