Team:Groningen/project characterisation mu lasr

From 2011.igem.org

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=LasR Autoinducing Loops=
=LasR Autoinducing Loops=
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Our second memory unit consists of a LasR gene(BBa_) driven the LasB promoter(BBa_). LasR only activates transcription in the presence of HSL, this was added to the medium in a concentration of 100nM.  
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Our second memory unit consists of a LasR gene(Bba_C0179) driven the LasB promoter(Bba_R0079). LasR only activates transcription in the presence of HSL, this was added to the medium in a concentration of 100nM.  
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==Leakiness of the LasR autoinducing loops==
{| class="wikitable"
{| class="wikitable"
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! colspan="2" | cI autoinducing loop 1 (BBa_K00) after 4 hours of incubation
 
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|-
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| [[File:ci1ui.png]]
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| || no-tag|| DAS || LVA ||   
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Figure 1. A dotplot of uninduced cells carrying the pSB1K3 plasmid with the biobrick BBa_ and the plasmid pSB1A3 with as well an input(PBAD controlling cI) as well as the output (PRM controlling GFP-LVA).
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|  
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{| class="wikitable"
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!  
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! Number
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! % of total
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! Mean fluorescence
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|-
|-
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| Visible
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| RBS 0,07 || <gallery>[[File:lasR9.png]] | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K607031 BBa_K607031] </gallery> || <gallery>[[File:lasR5.png]] | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K607027 BBa_K607027] </gallery>
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| 50000
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  || <gallery>[[File:lasR1.png]] | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K607023 BBa_K607023] </gallery>
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| 100
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|rowspan="4"| [[File:legenda.png]]
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| ---
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|-
|-
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| Fluorescent || 9180 || 18,36 || 11419
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| RBS 0,3 || <gallery>[[File:lasR10.png]]| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K607032 BBa_K607032]</gallery> || <gallery>[[File:lasR6.png]]| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K607028 BBa_K607028]</gallery> || <gallery>[[File:lasR2.png]]| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K607024 BBa_K607024]</gallery>
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|-
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| Non-Fluorescent|| 34424 || 68 || 5974
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| RBS 0,6 || <gallery> [[File:lasR11.png]]|[http://partsregistry.org/wiki/index.php?title=Part:BBa_K607033 BBa_K607033] </gallery>
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|}
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|| <gallery>[[File:lasR7.png]]| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K607029 BBa_K607029]</gallery> || <gallery>[[File:lasr3.png]]| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K607025 BBa_K607025]</gallery>
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|-
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| colspan=2 | Two populations are visible on the dotplot. We seperated both populations on the histogram to show that the major peak of nonfluorescent cells overlaps the minor peak of fluorescenting cells.
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| RBS 1 || <gallery>[[File:lasr12.png]]| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K607034 BBa_K607034]</gallery> || <gallery>[[File:lasr8.png]]| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K607030 BBa_K607030]</gallery> || <gallery>[[File:lasr4.png]]| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K607026 BBa_K607026]</gallery>
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|}
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{| class="wikitable"
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! colspan="2" | cI autoinducing loop 6 (BBa_K00) after 4 hours of incubation
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|-
|-
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| [[File:ci6ui.png]]
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|colspan="4"| '''Figure 4.''' Flow cytometer data taken at different time-points during exponential growth of cells carrying different cI autoinducing loops.
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Figure 2. A dotplot of uninduced cells carrying the pSB1K3 plasmid with the biobrick BBa_ and the plasmid pSB1A3 with as well an input(PBAD controlling cI) as well as the output (PRM controlling GFP-LVA).
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|  
|  
-
{| class="wikitable"
 
-
! 
 
-
! Number
 
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! % of total
 
-
! Mean fluorescence
 
-
|-
 
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| Visible
 
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| 50000
 
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| 100
 
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| ---
 
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|-
 
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| Fluorescent || 9180 || 18,36 || 11419
 
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|-
 
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| Non-Fluorescent|| 34424 || 68 || 5974
 
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|}
 
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|-
 
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| colspan=2 | Only one population is visible on the dotplot. The histogram was obtained in the same fashion as in figure 1 to show that there are cultures with activated cI loops and cultures without.
 
|}
|}
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{{FooterGroningen2011}}
{{FooterGroningen2011}}

Revision as of 22:11, 21 September 2011


LasR Autoinducing Loops

Our second memory unit consists of a LasR gene(Bba_C0179) driven the LasB promoter(Bba_R0079). LasR only activates transcription in the presence of HSL, this was added to the medium in a concentration of 100nM.

Leakiness of the LasR autoinducing loops

no-tag DAS LVA
RBS 0,07
Legenda.png
RBS 0,3
RBS 0,6
RBS 1
Figure 4. Flow cytometer data taken at different time-points during exponential growth of cells carrying different cI autoinducing loops.




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