Difference between revisions of "Part:BBa K118011:Experience"

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Our experiements was performed as bellow:
 
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<br>The correct clones were cultured in 96-well plate at 37℃ for 24 hours,then the fluorescence and absorbance at 600 nm were recorded on a SpectraMax M2 plate reader.All fluorescence was normalized with absorbance at 600 nm.The cell density was characterized by the absorbance at 600nm,which increased with glucose concentration.The result is showed in the figure bellow.
 
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<img src="https://static.igem.org/mediawiki/2012/a/a2/18D.png" width="496" height="428" hspace="2" vspace="1" border="2" align="center" />
 
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  In this figure, the fluorescence of RFP generator promoted by PcstA decreased when concentration of glucose rose,that is to say,promoter PcstA can be activated by CRP at low glucose concentration.
 
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<I>WHU-China 2012</I>
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We embedded the gene of RFP downstream the promoter PcstA. The correct clones were cultured in 96-well plate at 37℃ for 24 hours,then the fluorescence and absorbance at 600 nm were recorded on a SpectraMax M2 plate reader.All fluorescence was normalized with absorbance at 600 nm.The cell density was characterized by the absorbance at 600nm,which increased with glucose concentration.The result is showed in the figure bellow. 
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[[Image:https://static.igem.org/mediawiki/2012/a/a2/18D.png]]
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In this figure, the fluorescence of RFP generator promoted by PcstA decreased when concentration of glucose rose,that is to say,promoter PcstA can be activated by CRP at low glucose concentration. 
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Revision as of 17:03, 26 October 2012

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_K118011

Preliminary tests of this part were conducted using the reporter gene xylE (BBa_K118021). Strong repression occurred in the presence of glucose, and partial repression in the presence of high concentrations of higher sugars. Results can be found [http://2008.igem.org/Team:Edinbrugh/Results/PcstA-xylE here].

Characterization of PcstA with RFP Generator Part:BBa K081014

User Reviews

UNIQafd0b4c459b766b8-partinfo-00000001-QINU

BBa_K118011 1 Not understood Kun


II09 CRP-GFP fluor different media.jpg

Cells with BBa_K200018 were grown overnight in various different media, and the GFP fluorescence was measured.

After overnight culture, the corrected fluorescence of glucose is almost negligible, showing that glucose represses the PcstA promoter strongly.

For all the other secondary carbon sources, 10% Casamino Acids in M9 shows the highest corrected fluorescence at 22000.

For more information, go to the Imperial iGEM 2009 E.ncapsulator project page on [http://2009.igem.org/Team:Imperial_College_London/Wetlab/Results/CRP_and_Media BBa_K200018 testing results]

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UNIQafd0b4c459b766b8-partinfo-00000003-QINU

UNIQafd0b4c459b766b8-partinfo-00000004-QINU

BBa_K118011 2 Not understood WHU-China 2012

We embedded the gene of RFP downstream the promoter PcstA. The correct clones were cultured in 96-well plate at 37℃ for 24 hours,then the fluorescence and absorbance at 600 nm were recorded on a SpectraMax M2 plate reader.All fluorescence was normalized with absorbance at 600 nm.The cell density was characterized by the absorbance at 600nm,which increased with glucose concentration.The result is showed in the figure bellow.

File:Https://static.igem.org/mediawiki/2012/a/a2/18D.png

In this figure, the fluorescence of RFP generator promoted by PcstA decreased when concentration of glucose rose,that is to say,promoter PcstA can be activated by CRP at low glucose concentration.

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UNIQafd0b4c459b766b8-partinfo-00000006-QINU