Difference between revisions of "Part:BBa K4164015"
 
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The composite part can be directly imported into plasmid and express RXR-ddRFPA1 induced with IPTG.
 
The composite part can be directly imported into plasmid and express RXR-ddRFPA1 induced with IPTG.
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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.
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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 05:31, 12 October 2022


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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1964
    Illegal NheI site found at 2232
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 454
    Illegal AgeI site found at 1481
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 836