Difference between revisions of "Part:BBa K5301017"

 
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<partinfo>BBa_K5301017 short</partinfo>
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sGFP1-10 tether is composed of mSA, a 3C linker, a 6�His tag, and sGFP 1-10.We construct membrane protein dimers through the interaction between biotin and streptavidin, as well as the self-assembly mechanism of sGFP.
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===Usage and Biology===
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We devised the fusion expression of SdyCatcher with spNW15, SpyCatcher and mCherry[1-10] (namely SCSdC-mCh[1-10]). We employed SdyTag and SdyCatcher to link two proteins - SCSdC-mCh[1-10] and SnCSdT.
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===Characterization===
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We used SDS-PAGE to test whether the protein containing SdyCatcher had been expressed successfully(Figure 1). The molecular weight of SCSdC-mCh[1-10] (containing SdyCatcher) is 76.3 kDa. And we purified the target protein with a molecular weight of approximately 70-100 kD (Lane 4 - Lane 5), which demonstrated that we had successfully expressed the protein containing SdyCatcher.
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<div class="center"><div class="thumb tnone"><div class="thumbinner" style="width:min-content;"><div style="zoom:0.5;overflow:hidden;">
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https://static.igem.wiki/teams/5301/parts/sds-page-result-of-sdycatcher-for-sdycatcher.png
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Figure 1.SDS-PAGE analysis of the extraction results of SCSdC-mCh[1-10] (containing SdyCatcher).The gel was run at 80 V for 10 minutes and then at 150 V for 20 minutes, followed by staining with Coomassie Brilliant Blue. The molecular weight of SCSdC-mCh[1-10] is 76.3 kDa.
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K5301017 SequenceAndFeatures</partinfo>
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===Functional Parameters===
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<partinfo>BBa_K5301017 parameters</partinfo>
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Revision as of 17:50, 30 September 2024


sGFP1-10 tether is composed of mSA, a 3C linker, a 6His tag, and sGFP 1-10.

sGFP1-10 tether is composed of mSA, a 3C linker, a 6�His tag, and sGFP 1-10.We construct membrane protein dimers through the interaction between biotin and streptavidin, as well as the self-assembly mechanism of sGFP.

Usage and Biology

We devised the fusion expression of SdyCatcher with spNW15, SpyCatcher and mCherry[1-10] (namely SCSdC-mCh[1-10]). We employed SdyTag and SdyCatcher to link two proteins - SCSdC-mCh[1-10] and SnCSdT.

Characterization

We used SDS-PAGE to test whether the protein containing SdyCatcher had been expressed successfully(Figure 1). The molecular weight of SCSdC-mCh[1-10] (containing SdyCatcher) is 76.3 kDa. And we purified the target protein with a molecular weight of approximately 70-100 kD (Lane 4 - Lane 5), which demonstrated that we had successfully expressed the protein containing SdyCatcher.

sds-page-result-of-sdycatcher-for-sdycatcher.png

Figure 1.SDS-PAGE analysis of the extraction results of SCSdC-mCh[1-10] (containing SdyCatcher).The gel was run at 80 V for 10 minutes and then at 150 V for 20 minutes, followed by staining with Coomassie Brilliant Blue. The molecular weight of SCSdC-mCh[1-10] is 76.3 kDa.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 82
    Illegal AgeI site found at 142
  • 1000
    COMPATIBLE WITH RFC[1000]