Difference between revisions of "Part:BBa K5301014"
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<partinfo>BBa_K5301014 short</partinfo> | <partinfo>BBa_K5301014 short</partinfo> | ||
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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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| + | ==Usage and Biology== | ||
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| + | 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. | ||
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| + | ==Characterization== | ||
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| + | We performed an in vitro mixing and incubation of sGFP1-10 with sGFP11 to achieve their binding(in the form of sGFP1-10 tether <partinfo>BBa_K5301017</partinfo>and sGFP11 tether <partinfo>BBa_K5301018</partinfo>). The images captured under a fluorescence microscope are shown in the figure below. | ||
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| + | <div class="center"><div class="thumb tnone"><div class="thumbinner" style="width:min-content;"><div style="zoom:0.25;overflow:hidden;"> | ||
| + | https://static.igem.wiki/teams/5301/parts/sgfp-fluorescent.png | ||
| + | </div><div class="thumbcaption"> | ||
| + | Figure 1.Fluorescence images following the reassembly of sGFP(4×10).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. | ||
| + | </div></div></div></div> | ||
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| + | ==Sequence and Features== | ||
<partinfo>BBa_K5301014 SequenceAndFeatures</partinfo> | <partinfo>BBa_K5301014 SequenceAndFeatures</partinfo> | ||
Latest revision as of 08:56, 2 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(in the form of sGFP1-10 tether BBa_K5301017and sGFP11 tether BBa_K5301018). The images captured under a fluorescence microscope are shown in the figure below.
Figure 1.Fluorescence images following the reassembly of sGFP(4×10).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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]