Coding

Part:BBa_K4785005:Design

Designed by: ZhengLin Li   Group: iGEM23_SUSTech-CHINA   (2023-10-10)
Revision as of 06:39, 11 October 2023 by SeanHANPT (Talk | contribs) (→‎Design Notes)


high-mobility group box 1 (hmgb1)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 73


Design Notes

The protein expressed by this sequence is a nuclear protein, and the signal peptide sequence needs to be changed if it is expressed in the eukaryotic system or secreted out of the cell.

Figure 1: To purify the target proteins, we construct five plasmids: pET28a-6×His-HMGB1-FL, pET28a-6×His-HMGB1-AB, pET28a-6×His-HMGB1-A, pET28a-6×His-HMGB1-B, pET28a-6×His-PslG. After sequencing, we determine that the plasmid was correctly sequenced and ready for use.

Figure 2: The plasmids with the correct sequencing results were transferred into Escherichia coli BL21 for induced expression, then Escherichia coli was broken, and the target protein was purified by affinity chromatography and molecular sieve, and we obtained high purity of the target protein.

Source

Organism: Homo sapiens

References