Difference between revisions of "Part:BBa K838002"
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| − | Melanopsin is a photosensitive protein membrane | + | Melanopsin is a photosensitive protein membrane bound G protein coupled receptor. Photoactivation of a cis retinal (R) chromo- |
phore changes the comformation of melanopsin which in turn activates G alpha type G protein. The G alpha sets off a signaling cascade which eventually results in the activation of transient receptor potential channels which will allow calcium influx. The increased calcium concentration in the cell activates the calmodulin -> calcineurin -> NFAT pathway. The end result is the activation of genes situated next to NFAT promoter elements. | phore changes the comformation of melanopsin which in turn activates G alpha type G protein. The G alpha sets off a signaling cascade which eventually results in the activation of transient receptor potential channels which will allow calcium influx. The increased calcium concentration in the cell activates the calmodulin -> calcineurin -> NFAT pathway. The end result is the activation of genes situated next to NFAT promoter elements. | ||
Revision as of 19:50, 26 September 2012
Melanopsin
Melanopsin is a photosensitive protein membrane bound G protein coupled receptor. Photoactivation of a cis retinal (R) chromo- phore changes the comformation of melanopsin which in turn activates G alpha type G protein. The G alpha sets off a signaling cascade which eventually results in the activation of transient receptor potential channels which will allow calcium influx. The increased calcium concentration in the cell activates the calmodulin -> calcineurin -> NFAT pathway. The end result is the activation of genes situated next to NFAT promoter elements.
Just to recapitulate, this part is a membrane bound receptor. All other pathways used are endogenous.
==
Usage and Biology
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 282
Illegal PstI site found at 853
Illegal PstI site found at 1311 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 282
Illegal PstI site found at 853
Illegal PstI site found at 1311 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1369
Illegal BamHI site found at 45
Illegal BamHI site found at 541 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 282
Illegal PstI site found at 853
Illegal PstI site found at 1311 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 282
Illegal PstI site found at 853
Illegal PstI site found at 1311 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 711
How to use
1) Obtain the parts: To implement a light activated gene switch using this protein, all one needs is this part and any gene to express in a mammalian cell.
2) Cloning:
First clone the part into a mammalian expression vector such as [http://products.invitrogen.com/ivgn/product/V79020?ICID=search-product pcDNA3.1(+)] or [http://products.invitrogen.com/ivgn/product/V04450 pCEP4]. These are the two main expression vectors we used. To use the melanopsin pathway, you also need to clone any gene you want to express into the mammalian vector [http://www.promega.com/resources/protocols/product-information-sheets/a/pgl430-vector-protocol/ pGL4.30].
3) Transfection
Co-transfect the combination of photoreceptive protein and readout construct of your choice in mammalian cells!
