Team:LMU-Munich/Jump-or-die/Functional Principle
From 2010.igem.org
(Difference between revisions)
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Construct 1 contains a tet-on promoter, bacterial attachment site (attB), human bak (bak) and SV40 polyadenylation site (SV40PA). It has to be integrated to the genom of the cellline. The selection if this construct is integrated will be made by double transfection with hygromycin resistance and tet-on-inducible apoptosis. | Construct 1 contains a tet-on promoter, bacterial attachment site (attB), human bak (bak) and SV40 polyadenylation site (SV40PA). It has to be integrated to the genom of the cellline. The selection if this construct is integrated will be made by double transfection with hygromycin resistance and tet-on-inducible apoptosis. | ||
- | [[Image: | + | [[Image: Jump11.jpg | 400px | Jump construct 1]] |
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Construct 2 will be inserted in a vector and consist of the following parts: phage attachment site (attP), eGFP (our gene of interest), doubled stop codon and SV40PA. | Construct 2 will be inserted in a vector and consist of the following parts: phage attachment site (attP), eGFP (our gene of interest), doubled stop codon and SV40PA. | ||
- | [[Image: | + | [[Image: Jump12.jpg | 400px | Jump construct 2]] |
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This Biobrick is containing CMV-promoter, PhiC31o (mouse-codon optimized integrase) and SV40PA. It will be transfected to the cellline and read-off so that PhiC31o will be existing. | This Biobrick is containing CMV-promoter, PhiC31o (mouse-codon optimized integrase) and SV40PA. It will be transfected to the cellline and read-off so that PhiC31o will be existing. | ||
- | [[Image: | + | [[Image: Jump13.jpg | 400px | Jump construct 3]] |
== Transfection possibilities == | == Transfection possibilities == | ||
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PhiC31o will be read off because of CMV promoter. Now the integrase will combine attB and attP site and integrate construct 2 into genom. Up next, the gene of interest is behind tet-on promoter and will be read off after induction. The doubled stop codon will efficiently prevent the tranlation of bak. This cell will express the gene of interest and survive. | PhiC31o will be read off because of CMV promoter. Now the integrase will combine attB and attP site and integrate construct 2 into genom. Up next, the gene of interest is behind tet-on promoter and will be read off after induction. The doubled stop codon will efficiently prevent the tranlation of bak. This cell will express the gene of interest and survive. | ||
+ | [[Image: Jump14.jpg | 400px | Jump recombination]] | ||
{{:Team:LMU-Munich/Templates/Page Footer}} | {{:Team:LMU-Munich/Templates/Page Footer}} |
Revision as of 12:09, 18 August 2010