Abstract：Objective: To express and purify human HMGB1 A box fusion protein, PTD-mLumin-HMGB1 A, which contains a protein transduction domain and a newly isolated far-red fluorescent protein named mLumin, and investigate its ability to across the membrane. Methods: Expression vector of PTD-mLumin-HMGB1 Awas constructed with inserting the HMGB1 A box into pET28a -PTD-mLumin vector. The fusion protein was expressed by E.coli Rossetta (DE3) and purified by Nicolumn. Confocal microscopy was used to confirm the transmembrane ability ofPTD-mLumin-HMGB1 A. Results: The vector of pET28a-PTD-mLumin-HMGB1 A was successfully constructed, and the fusion protein expressed and was purified efficiently.PTD-mLumin-HMGB1 Afusion protein transduced into EL-4 cell efficiently. Conclusion: The PTD-mLumin-HMGB1 Aprotein was successfully constructed, which laid the foundation for further investigation of biological function of human HMGB1 A box.
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2012, Vol. 22 
