Difference between revisions of "Part:BBa K5301014"
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sGFP11 is the smaller fragment of the Split-GFP system, encompassing the final β-strand domain of green fluorescent protein (GFP).The isolated GFP11 fragment does not fluoresce on its own and is typically fused to the C-terminus of the target protein. Upon binding with the GFP1-10 fragment, it facilitates the restoration of GFP's fluorescent activity.
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sGFP11 is the smaller fragment of the Split-GFP system, encompassing the final β-strand domain of green fluorescent protein (GFP).The isolated GFP11 fragment does not fluoresce on its own and is typically fused to the C-terminus of the target protein. Upon binding with the GFP1-10 fragment(<partinfo>BBa_K5301012</partinfo>), it facilitates the restoration of GFP's fluorescent activity.
  
  
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Figure 1.Fluorescence images following the reassembly of sGFP(4×).Figure A and Figure C represent the background fluorescence, while Figures B and D show the observations after the addition of the samples at corresponding positions. By comparing these results, the corresponding fluorescence signals are obtained to eliminate false positives.
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Figure 1.Fluorescence images following the reassembly of sGFP(40×).Figure A and Figure C represent the background fluorescence, while Figures B and D show the observations after the addition of the samples at corresponding positions. By comparing these results, the corresponding fluorescence signals are obtained to eliminate false positives.
 
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Revision as of 15:04, 1 October 2024


sGFP11 is an engineered version of the final strand of the GFP beta-barrel.


sGFP11 is the smaller fragment of the Split-GFP system, encompassing the final β-strand domain of green fluorescent protein (GFP).The isolated GFP11 fragment does not fluoresce on its own and is typically fused to the C-terminus of the target protein. Upon binding with the GFP1-10 fragment(BBa_K5301012), it facilitates the restoration of GFP's fluorescent activity.


Usage and Biology

In our team's experiments, sGFP1-10 and sGFP11 are individually fused and expressed with the target proteins.We use the self-assembly of GFP1-10 and GFP11 to construct membrane protein dimers.


Characterization

We performed an in vitro mixing and incubation of sGFP1-10 with sGFP11 to achieve their binding. The images captured under a fluorescence microscope are shown in the figure below.


sgfp-fluorescent.png

Figure 1.Fluorescence images following the reassembly of sGFP(40×).Figure A and Figure C represent the background fluorescence, while Figures B and D show the observations after the addition of the samples at corresponding positions. By comparing these results, the corresponding fluorescence signals are obtained to eliminate false positives.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]