Difference between revisions of "Part:BBa K257023:Experience"

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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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===Applications of BBa_K257023===
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===Caracterisation of BBa_K257023===
  
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[[Image:PFec-mRFP1.JPG|Pfec caracterisation]]
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Overnight cultures of BBa_K257023 were diluted 1/100 in LB or M9 medium supplemented with varing amont of FeNO3 and Na Citrate. Fluorescence and OD were assessed in a Tecan plate reader. Fluorescence divided by OD is ploted.
  
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Caracterisation of Fec promoter upon induction with increasing Na Citrate concentrations, on a growth medium with varaiable concentrations of Fe(NO3)3
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[[Image:PFec-mRFP1-LB.JPG|Pfec caracterisation|500px]]
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[[Image:PFec-mRFP1-M9.JPG|Pfec caracterisation|500px]]
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The low Fluo/OD at high FeNO3 concentration can be explained by the taxicity of FeNO3. Note that the best response to Na Citrate is obtained with 1mM FeNO3.
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=== Induction===
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pFec-RFP on LB medium
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[[Image:LB.jpg]]
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pFec-RFP on medium with Na citrate (30mM)
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[[Image:citrate30.jpg]]
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pFec-RFP on medium with Fe citrate (30mM)
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[[Image:citrate30 Fe1.jpg]]
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We've got a control on LB medium : fluorescence is weak. With a concentration of 30mM (Na citrate or Fe citrate), we have the same level of induction of pFec. We can explain this because of the relative toxicity of the Fe(NO3)3 we use to get Fe citrate. Besides, for this amount of citrate, we can expect Fe to be entirely chelated by the citrate. Nevertheless, we demonstrate the induction of Fec promoter.
  
 
===User Reviews===
 
===User Reviews===

Latest revision as of 01:06, 22 October 2009


Caracterisation of BBa_K257023

Overnight cultures of BBa_K257023 were diluted 1/100 in LB or M9 medium supplemented with varing amont of FeNO3 and Na Citrate. Fluorescence and OD were assessed in a Tecan plate reader. Fluorescence divided by OD is ploted.

Pfec caracterisation Pfec caracterisation

The low Fluo/OD at high FeNO3 concentration can be explained by the taxicity of FeNO3. Note that the best response to Na Citrate is obtained with 1mM FeNO3.

Induction

pFec-RFP on LB medium

LB.jpg


pFec-RFP on medium with Na citrate (30mM)

Citrate30.jpg


pFec-RFP on medium with Fe citrate (30mM)

Citrate30 Fe1.jpg

We've got a control on LB medium : fluorescence is weak. With a concentration of 30mM (Na citrate or Fe citrate), we have the same level of induction of pFec. We can explain this because of the relative toxicity of the Fe(NO3)3 we use to get Fe citrate. Besides, for this amount of citrate, we can expect Fe to be entirely chelated by the citrate. Nevertheless, we demonstrate the induction of Fec promoter.

User Reviews

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