Difference between revisions of "Part:BBa J23117:Experience"

Revision as of 14:39, 27 October 2013

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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

UNIQe6d5bd7383ddb4be-partinfo-00000000-QINU UNIQe6d5bd7383ddb4be-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

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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.
Our results from these approaches showed that BBa_J23117 has the lowest promoter activity compared to BBa_J23116, BBa_J23110 and BBa_J23118.

Fig. 1. RFP and qRT-PCR promoter analyses.

Fig. 2. Microscopic promoter analyses on single cell level
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

@@ Line 47: / Line 47: @@
 The promoter strength was measured by using the reporter gene ''rfp''.<br />
-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.<br />
+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.<br />
 Our results from these approaches showed that [[Part:BBa_J23117|BBa_J23117]] has the lowest promoter activity compared to [[Part:BBa_J23116|BBa_J23116]], [[Part:BBa_J23110|BBa_J23110]] and [[Part:BBa_J23118|BBa_J23118]].