Part:BBa_K4164015
Inductive expression of RXR-ddRFPB1
In order to find an appropriate expression intensity to achieve balance between metabolic burden and detection efficiency, we also tried T7lac promoter from pET-29a+(Novagen).
The composite part can be directly imported into plasmid and express RXR-ddRFPA1 induced with IPTG.
We tried to determine the solubility of RXR-ddRFPB1 by purifying 6xHis-tagged proteins under native conditions and performing SDS-PAGE to compare the abundance of the band with the size of interest.As shown in the Figure 1, RXR-ddRFPB1(78.9kDa) is soluble and can be purified by Ni-NTA affinity chromatography.
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]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1964
Illegal NheI site found at 2232 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 454
Illegal AgeI site found at 1481 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 836
None |