Team:EPF-Lausanne/Parts

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First clone the part into a mammalian expression vector such as pcDNA3.1(+)[http://products.invitrogen.com/ivgn/product/V79020?ICID=search-product] or pCEP4 [http://products.invitrogen.com/ivgn/product/V04450]. These are the two main expression vectors we used.
First clone the part into a mammalian expression vector such as pcDNA3.1(+)[http://products.invitrogen.com/ivgn/product/V79020?ICID=search-product] or pCEP4 [http://products.invitrogen.com/ivgn/product/V04450]. These are the two main expression vectors we used.
To use the melanopsin pathway, you also need to clone a target gene of your choice downstream pNFAT.
To use the melanopsin pathway, you also need to clone a target gene of your choice downstream pNFAT.
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[[File:Team_EPF_Lausanne_pNFAT.png|100px]] -> [[File:Team_EPF_Lausanne_pNFAT+target.png|300px]]
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*[[File:Team_EPF_Lausanne_pNFAT.png|100px]] -> [[File:Team_EPF_Lausanne_pNFAT+target.png|300px]]
'''2) Choose your switch:'''
'''2) Choose your switch:'''
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Simple switch = [[File:Team_EPF_Lausanne_LovTAP.png|300px]] + [[File:Team_EPF_Lausanne_RO.png|300px]]
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*Simple switch = [[File:Team_EPF_Lausanne_LovTAP.png|300px]] + [[File:Team_EPF_Lausanne_RO.png|300px]]
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Complex switch = [[File:Team_EPF_Lausanne_melanopsin.png|200px]] + [[File:Team_EPF_Lausanne_pNFAT+target.png|300px]]
+
*Complex switch = [[File:Team_EPF_Lausanne_melanopsin.png|200px]] + [[File:Team_EPF_Lausanne_pNFAT+target.png|300px]]
3) Transfection
3) Transfection

Revision as of 00:06, 24 September 2012

Our parts

We managed to submit 4 new parts that will give other teams the possibility to reproduce our experiments with both the switches we tested.

The melanopsin coding gene and the NFAT response element will allow to use the more complex switch, by just cloning melanopsin into a mammalian vector and clone a readout gene (fluorescent protein, quantitatively measurable protein) downstream NFAT and clone both in a mammalian vector.

We also submitted our favorite parts, which are a mammalian LovTAP-VP16 and a composite readout system build to respond to it. Both parts only need cloning into a mammalian vector to be ready for transfection.

How to use

1) Cloning: First clone the part into a mammalian expression vector such as pcDNA3.1(+)[http://products.invitrogen.com/ivgn/product/V79020?ICID=search-product] or pCEP4 [http://products.invitrogen.com/ivgn/product/V04450]. These are the two main expression vectors we used. To use the melanopsin pathway, you also need to clone a target gene of your choice downstream pNFAT.

  • Team EPF Lausanne pNFAT.png -> Team EPF Lausanne pNFAT+target.png

2) Choose your switch:

  • Simple switch = Team EPF Lausanne LovTAP.png + Team EPF Lausanne RO.png
  • Complex switch = Team EPF Lausanne melanopsin.png + Team EPF Lausanne pNFAT+target.png

3) Transfection Co-transfect the combination of photoreceptive protein and readout construct of your choice in mammalian cells!



<groupparts>iGEM012 EPF-Lausanne</groupparts>

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