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Team:Slovenia/PROJECT/proof/popfret/fret
From 2010.igem.org
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| - | + | <p><span style="font-family: Tahoma;" lang="en-gb" xml:lang="en-gb">To prove that DNA program sequence could serve as a scaffold for more than two chimeric proteins, four split GFPs linked to zinc fingers were used. We predicted that DNA program sequence will promote a reconstitution of both pairs of split GFPs and FRET among reconstituted split GFPs. To achive that, cells have to express all four chimeric proteins and contain a DNA program sequence as well, all localised in the same region. Proving the principle in this way was achieved by FRET in mammalian cells.</span><span style="font-family: Tahoma; line-height: 15px;"> Selection of cells for FRET measurements is critical and is demonstrated below as well as proof for nuclear colocalisation of reconstituted split GFPs.</span></p> | |
Revision as of 16:54, 27 October 2010
To prove that DNA program sequence could serve as a scaffold for more than two chimeric proteins, four split GFPs linked to zinc fingers were used. We predicted that DNA program sequence will promote a reconstitution of both pairs of split GFPs and FRET among reconstituted split GFPs. To achive that, cells have to express all four chimeric proteins and contain a DNA program sequence as well, all localised in the same region. Proving the principle in this way was achieved by FRET in mammalian cells. Selection of cells for FRET measurements is critical and is demonstrated below as well as proof for nuclear colocalisation of reconstituted split GFPs.
