Team:Evry/Tadpole injection1

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Line 23: Line 23:
<h2><li>pCS2+ GFP-aid</li> </h2><br/>
<h2><li>pCS2+ GFP-aid</li> </h2><br/>
<h3>24h after injection</h3><br/>
<h3>24h after injection</h3><br/>
 +
Eggs are near stages 20, neural crest is clearly visible <br/>
<img src="/wiki/images/c/c7/409GFP-aid%2Bcontrol.jpg" alt="perdu" /><br/>
<img src="/wiki/images/c/c7/409GFP-aid%2Bcontrol.jpg" alt="perdu" /><br/>

Revision as of 01:40, 18 September 2012

Plasmid injection in fertilized Xenopus laevis eggs


Plasmids injected:



We injected 2.3 nl of plasmid at 100ng.µl-1

  1. pCS2+ GFP-aid: contains the constitutive promoter CMV and the aid sequenced of the aid system fusionned to GFP (Green Fluorescent Protein)(Nishimura et al., 2009).
    2. Number of Plasmids injected: ~ 3.78E+7

  2. pCS2+ citrine: contains the constitutive promoter CMV and the fluorescent protein citrin (yellow).
    3. Number of Plasmids injected: ~ 4.38E+7

  3. pCS2+ mCFP: contains the constitutive promoter CMV and the fluorescent protein mCFP (Cyan Fluorescent Protein).
    4. Number of Plasmids injected: ~ 4.37E+7

  4. pCS2+ GFP-aid co-injected with the plasmid pNHK60 (from Nishimura et al., 2009) : contains the constitutive promoter CMV and the aid sequenced of the aid system fusionned to GFP, pNHK60 contains the CMV promoter followed by the TIR1 protein which in presence of auxin degrade form a complex with aid of GFP-aid and degrade it AID system.
    5. Number of Plasmids injected: ~ 1.89E+7 for pCS2+ GFP-aid and ~ 1.3E+7 for pNHK60

  1. pCS2+ GFP-aid


    2. 24h after injection


    Eggs are near stages 20, neural crest is clearly visible
    perdu
    perdu

  2. pCS2+ citrine



  3. pCS2+ mCFP



  4. pCS2+ GFP-aid co-injected with the plasmid pNHK60



References:


Nishimura, K. et al., 2009. An auxin-based degron system for the rapid depletion of proteins in nonplant cells. Nature Methods, 6(12), p.917-922.

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