Difference between revisions of "Part:BBa K4164005"
 
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<partinfo>BBa_K4164005 short</partinfo>
 
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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.
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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 ddRFP-A1. ddRFP-A1 interacts with ddRFP-B1 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. ddRFP-B1 does not form a chromatography group and exhibits no fluorescence in the monomeric state.
 
    
 
    
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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 Ca<sup>2+</sup>-dependent association of calmodulin, M13 in live cells, and imaging of caspase-3 activity during apoptosis.
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ddRFP-A1 can form a heterodimer with ddRFP-B1, 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 Ca<sup>2+</sup>-dependent association of calmodulin, M13 in live cells, and imaging of caspase-3 activity during apoptosis.
  
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We connected ddRFP-A1 and ddRFP-B1 by a flexible linker(3*GGGGS) to verify the feasicility of ddRFP-A1 and ddRFP-B1. Following overnight incubation at 37℃, we can see the bright red fluorescence (Fig.1). In this case, we chose ddRFP-A1 and ddRFP-B1 as our report device.
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<p style="text-align: center;"><img src="https://static.igem.wiki/teams/4164/wiki/part-registry/part-004005016998ddrfpplate.png"with="1000" height="" width="500" height=""/></p>
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<p style="text-align: center!important;"><b>Fig. 1. Fluorescence image of <em> E. coli</em> expressing ddRFPA1-ddRFPB1 and control.</b></p>
 
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<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Latest revision as of 14:13, 13 October 2022


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 ddRFP-A1. ddRFP-A1 interacts with ddRFP-B1 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. ddRFP-B1 does not form a chromatography group and exhibits no fluorescence in the monomeric state.

ddRFP-A1 can form a heterodimer with ddRFP-B1, 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 ddRFP-A1 and ddRFP-B1 by a flexible linker(3*GGGGS) to verify the feasicility of ddRFP-A1 and ddRFP-B1. Following overnight incubation at 37℃, we can see the bright red fluorescence (Fig.1). In this case, we chose ddRFP-A1 and ddRFP-B1 as our report device.


Fig. 1. Fluorescence image of E. coli expressing ddRFPA1-ddRFPB1 and control.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 436
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]