Difference between revisions of "Part:BBa K4164004"
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The dimerization-dependent red fluorescent protein-A1 (ddRFP-A1) is a variant of the dimeric Tomato, which is a monomer protein of the red fluorescent protein heterodimer, with a size of 26.4kDa. | The dimerization-dependent red fluorescent protein-A1 (ddRFP-A1) is a variant of the dimeric Tomato, which is a monomer protein of the red fluorescent protein heterodimer, with a size of 26.4kDa. | ||
| − | ddRFP-A1 is an RFP variant with weak fluorescence derived from the directed evolution of dTomato gene. It has a chromogenic group and can exhibit weak fluorescence in the monomeric state. ddRFPA1 can form a heterodimer with ddRFPB1, increasing the fluorescence intensity ten times more than the dissociation state. The utility of ddRFP-A1B1 has been demonstrated in three applications, including the detection of a protein-protein interaction in vitro, imaging of the reversible | + | ddRFP-A1 is an RFP variant with weak fluorescence derived from the directed evolution of dTomato gene. It has a chromogenic group and can exhibit weak fluorescence in the monomeric state. ddRFPA1 can form a heterodimer with ddRFPB1, increasing the fluorescence intensity ten times more than the dissociation state. The utility of ddRFP-A1B1 has been demonstrated in three applications, including the detection of a protein-protein interaction in vitro, imaging of the reversible Ca<sub>2+</sub>-dependent association of calmodulin and imaging of caspase-3 activity during apoptosis. |
We connected ddRFPA1 and ddRFPB1 by a flexible linker(3*GGGGS) to verify the feasicility of ddRFPA1 and ddRFPB1. Following overnight incubation at 37℃, we can see the bright red fluorescence (Figure 1). In this case, we chose ddRFPA1 and ddRFPB1 as our report device. | We connected ddRFPA1 and ddRFPB1 by a flexible linker(3*GGGGS) to verify the feasicility of ddRFPA1 and ddRFPB1. Following overnight incubation at 37℃, we can see the bright red fluorescence (Figure 1). In this case, we chose ddRFPA1 and ddRFPB1 as our report device. | ||
Revision as of 07:02, 12 October 2022
dimerization-dependent red fluorescent protein-A1(ddRFP-A1)
The dimerization-dependent red fluorescent protein-A1 (ddRFP-A1) is a variant of the dimeric Tomato, which is a monomer protein of the red fluorescent protein heterodimer, with a size of 26.4kDa.
ddRFP-A1 is an RFP variant with weak fluorescence derived from the directed evolution of dTomato gene. It has a chromogenic group and can exhibit weak fluorescence in the monomeric state. ddRFPA1 can form a heterodimer with ddRFPB1, increasing the fluorescence intensity ten times more than the dissociation state. The utility of ddRFP-A1B1 has been demonstrated in three applications, including the detection of a protein-protein interaction in vitro, imaging of the reversible Ca2+-dependent association of calmodulin and imaging of caspase-3 activity during apoptosis.
We connected ddRFPA1 and ddRFPB1 by a flexible linker(3*GGGGS) to verify the feasicility of ddRFPA1 and ddRFPB1. Following overnight incubation at 37℃, we can see the bright red fluorescence (Figure 1). In this case, we chose ddRFPA1 and ddRFPB1 as our report device.
Fig.1 FXR+CDCA.
In our project, we fused ddRFP-A1 and ddRFP-B1 to the C-terminus of FXR and RXR, respectively. Taking advantage of the characteristics that RXR can bind to the FXR as a heterodimer, using FXR as the receptor, and when the downstream ddRFP-A1 and ddRFP-B1 are correspondingly driven to dimerize and exhibit red fluorescence.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]