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. | ||
| + | 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. | ||
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1468
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 1057