Difference between revisions of "Part:BBa K606026:Experience"

 
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===Applications and characterisation of BBa_K606026 - TetO Array===
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We characterize it with arabinose-induced YFP:tetR Wild Type from pFX234 plasmid of F-X barre and D. Lane (Kinetics of plasmid segregation, Molecular Microbiology, 2004).<br>
  
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__NOTOC__
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In order to do this characterization, we took pictures of different plasmids containing only TetO array (K606026); TetR + YFP (pFX234); TetO + TetR + YFP (K606026 and pFX234). In each case we made a control by non inducing the promoter with arabinose.
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
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how you used this part and how it worked out.
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<center>
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{| border="1" class="wikitable" style="text-align: center;" align="center"
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|+ tetO array : 37°C
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|-
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|[[File:teto_minus_trans.jpg|350px|thumb|center|tetO / tetO array inducted with no arabinose on E. Coli .]]
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|[[File:teto_minus_Fluo20.jpg|350px|thumb|center|tetO / tetO array inducted with no arabinose on E. Coli .]]
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|-
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|[[File:teto_Arab_trans.jpg|350px|thumb|center|tetO / tetO array inducted with 0,2% arabinose on E. Coli .]]
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|[[File:teto_arab_Fluo20.jpg|350px|thumb|center|tetO / tetO array inducted with 0,2% arabinose on E. Coli .]]
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|}
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{| border="1" class="wikitable" style="text-align: center;"
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|+tetR:YFP : 37°C
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|-
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|[[File:yfp_tetr_minus_trans.jpg|350px|thumb|center|tetR:YFP / tetr-YFP inducted with no arabinose on E. Coli .]]
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|[[File:yfp_tetr_minus_fluo20.jpg|350px|thumb|center|tetR:YFP / tetr-YFP inducted with no arabinose on E. Coli .]]
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|-
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|[[File:yfp_tetr_Arab_trans.jpg|350px|thumb|center|tetR:YFP / tetr-YFP inducted with 0,2% arabinose on E. Coli .]]
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|[[File:yfp_tetr_Arab_fluo20.jpg|350px|thumb|center|tetR:YFP / tetr-YFP inducted with 0,2% arabinose on E. Coli .]]
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|}
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{| border="1" class="wikitable" style="text-align: center;"
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|+tetR:YFP / tetO array : 37°C
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|-
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|[[File:yfp_tetr_teto_minus_trans.jpg|350px|thumb|center|tetR:YFP-tetO/ full construct inducted with no arabinose on E. Coli .]]
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|[[File:yfp_tetr_teto_minus_fluo20_2s.jpg|350px|thumb|center|tetR:YFP-tetO/ full construct inducted with no arabinose on E. Coli .]]
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|-
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|[[File:yfp_tetr_teto_Arab_trans_2.jpg|350px|thumb|center|tetR:YFP-tetO / full construct inducted with 0,2% arabinose on E. Coli .]]
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|[[File:yfp_tetr_teto_Arab_fluo20_2-1.jpg|350px|thumb|center|tetR:YFP-tetO / full construct inducted with 0,2% arabinose on E. Coli .]]
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|}
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</center>
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The pictures of tetO show no YFP activity, which is normal because there is no YFP sequence in these plasmids.<br>
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The tetR-YFP construct which constitutes the transmitter part, occasionally shows gross aggregated YFP. This is not what we expected at first, but that does not prevent us to characterize the full construct.<br>
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After observing the full construct's pictures, we can obviously distinguish glowing dots in some cells. They reflect the behavior we expected. Indeed, appearance of dots shows that the receiver (tetO array) actually links tightly to tetR-YFP which is the emitted protein. <br><br>
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More information on : [http://2011.igem.org/Team:Paris_Bettencourt/Experiments/YFP_TetR_diffusion]
  
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===Applications of BBa_K606026===
 
  
 
===User Reviews===
 
===User Reviews===

Revision as of 12:28, 19 September 2011

Applications and characterisation of BBa_K606026 - TetO Array

We characterize it with arabinose-induced YFP:tetR Wild Type from pFX234 plasmid of F-X barre and D. Lane (Kinetics of plasmid segregation, Molecular Microbiology, 2004).

In order to do this characterization, we took pictures of different plasmids containing only TetO array (K606026); TetR + YFP (pFX234); TetO + TetR + YFP (K606026 and pFX234). In each case we made a control by non inducing the promoter with arabinose.

tetO array : 37°C
tetO / tetO array inducted with no arabinose on E. Coli .
tetO / tetO array inducted with no arabinose on E. Coli .
tetO / tetO array inducted with 0,2% arabinose on E. Coli .
tetO / tetO array inducted with 0,2% arabinose on E. Coli .
tetR:YFP : 37°C
tetR:YFP / tetr-YFP inducted with no arabinose on E. Coli .
tetR:YFP / tetr-YFP inducted with no arabinose on E. Coli .
tetR:YFP / tetr-YFP inducted with 0,2% arabinose on E. Coli .
tetR:YFP / tetr-YFP inducted with 0,2% arabinose on E. Coli .
tetR:YFP / tetO array : 37°C
tetR:YFP-tetO/ full construct inducted with no arabinose on E. Coli .
tetR:YFP-tetO/ full construct inducted with no arabinose on E. Coli .
tetR:YFP-tetO / full construct inducted with 0,2% arabinose on E. Coli .
tetR:YFP-tetO / full construct inducted with 0,2% arabinose on E. Coli .

The pictures of tetO show no YFP activity, which is normal because there is no YFP sequence in these plasmids.
The tetR-YFP construct which constitutes the transmitter part, occasionally shows gross aggregated YFP. This is not what we expected at first, but that does not prevent us to characterize the full construct.
After observing the full construct's pictures, we can obviously distinguish glowing dots in some cells. They reflect the behavior we expected. Indeed, appearance of dots shows that the receiver (tetO array) actually links tightly to tetR-YFP which is the emitted protein.

More information on : [http://2011.igem.org/Team:Paris_Bettencourt/Experiments/YFP_TetR_diffusion]


User Reviews

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