Difference between revisions of "Part:BBa K4164994"
 
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This part includes the ddRFPA1 fused by a 3*GS Linker to the FXR, which can form a heterodimer RXR-3*GS Linker-ddRFPB1 and exhibit red fluorescence when activated by CDCA.
 
This part includes the ddRFPA1 fused by a 3*GS Linker to the FXR, which can form a heterodimer RXR-3*GS Linker-ddRFPB1 and exhibit red fluorescence when activated by CDCA.
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We tried to determine the solubility of FXR-ddRFPA1 by purifying 6xHis-tagged proteins under native conditions and performing SDS-PAGE to compare the abundance of the band with the size of interest.Initially, FXR-ddRFPA1 is in the pellet after lysis and can't be purified(Lane1), then we optimized the expression conditions and purified it from supernatant of bacteria liquid.
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<p style="text-align: center;"><img src="https://static.igem.wiki/teams/4164/wiki/part-registry/sds-page.png"with="1000" height="" width="500" height=""/>
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<p style="text-align: center;"><b>Fig.1 Purification of FXR-ddRFPA1 and RXR-ddRFPB1.Lane 1: protein contained in the pellet after bacterial disruption. Lane 2: RXR-ddRFPB1 purified from supernatant of bacteria liquid. Lane 3: FXR-ddRFPA1 purified from supernatant of bacteria liquid in optimized expression conditon..</b></p>
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 05:22, 12 October 2022


FXR-3*GS Linker-ddRFPA1

This part includes the ddRFPA1 fused by a 3*GS Linker to the FXR, which can form a heterodimer RXR-3*GS Linker-ddRFPB1 and exhibit red fluorescence when activated by CDCA.

We tried to determine the solubility of FXR-ddRFPA1 by purifying 6xHis-tagged proteins under native conditions and performing SDS-PAGE to compare the abundance of the band with the size of interest.Initially, FXR-ddRFPA1 is in the pellet after lysis and can't be purified(Lane1), then we optimized the expression conditions and purified it from supernatant of bacteria liquid.


Fig.1 Purification of FXR-ddRFPA1 and RXR-ddRFPB1.Lane 1: protein contained in the pellet after bacterial disruption. Lane 2: RXR-ddRFPB1 purified from supernatant of bacteria liquid. Lane 3: FXR-ddRFPA1 purified from supernatant of bacteria liquid in optimized expression conditon..


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1468
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1057