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Team:Evry/Tadpole injection1
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<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 near stage 20, neural fold is visible <br/> | + | Eggs are near stage 20, neural fold is 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/> | ||
<img src="/wiki/images/2/2e/409gfpaid2.jpg" alt="perdu" /><br/> | <img src="/wiki/images/2/2e/409gfpaid2.jpg" alt="perdu" /><br/> | ||
| - | Some eggs expressed GFP-aid and some others not, it means that the injection did not work or the eggs do not express GFP-aid | + | |
| + | Some eggs expressed GFP-aid and some others not, it means that the injection did not work for some eggs or the eggs do not express GFP-aid<br/> | ||
| + | |||
| + | <h3>48h after injection</h3><br/> | ||
| + | Embryos are at stage 34-38, | ||
<h2><li>pCS2+ citrine</li> </h2><br/><br/> | <h2><li>pCS2+ citrine</li> </h2><br/><br/> | ||
Revision as of 09:45, 18 September 2012
Plasmid injection in fertilized Xenopus laevis eggs
Plasmids injected:
We injected 2.3 nl of plasmid at 100ng.µl-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). Number of Plasmids injected: ~ 3.78E+7
- pCS2+ citrine: contains the constitutive promoter CMV and the fluorescent protein citrin (yellow). Number of Plasmids injected: ~ 4.38E+7
- pCS2+ mCFP: contains the constitutive promoter CMV and the fluorescent protein mCFP (Cyan Fluorescent Protein). Number of Plasmids injected: ~ 4.37E+7
- 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. Number of Plasmids injected: ~ 1.89E+7 for pCS2+ GFP-aid and ~ 1.3E+7 for pNHK60
- pCS2+ GFP-aid
- pCS2+ citrine
- pCS2+ mCFP
- pCS2+ GFP-aid co-injected with the plasmid pNHK60
24h after injection
Eggs are near stage 20, neural fold is visible
Some eggs expressed GFP-aid and some others not, it means that the injection did not work for some eggs or the eggs do not express GFP-aid
48h after injection
Embryos are at stage 34-38,
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.
