Part:BBa_K4164005
dimerization-dependent red fluorescent protein-B1(ddRFP-B1)
The dimerization-dependent red fluorescent protein-B1(ddRFP-B1) is a variant of the dimeric Tomato. DdRFP-B1 is a monomer protein of the red fluorescent protein heterodimer, with a size of 26.4kDa, which is a dTomato-derived partner capable of forming dimers with ddRFPA1. ddRFPA1 interacts with ddRFPB1 with a Kd of 33 μM, and, therefore,the heterodimers can exist primarily in a free state at cytoplasmic concentrations sufficient for live cell imaging. ddRFPB1 does not form a chromatography group and exhibits no fluorescence in the monomeric state.
dRFPA1 can form a heterodimer with ddRFPB1, increasing the fluorescence intensity ten times more than the dissociation state.The use 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, M13 in live cells, 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 (Fig.1). In this case, we chose ddRFPA1 and ddRFPB1 as our report device.
Fig.1 Fluorescence image of E. coli expressing ddRFPA1-ddRFPB1 and control. .
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 436
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
| None |