Objective     To design and prepare recombinant mutant human high mobility group box 1 protein (HMGB1s) that can combine with HMGB1 receptors but cannot produce inflammatory effect

and accordingly lead to the creation of a new potential agent for anti-inflammatory therapy.       Methods     This experiment was based on successful clone and expression of human HMGB1.Six mutant  HMGB1 cDNA were designed and constructed by one step inverse PCR. They were cloned into prokaryotic expressive vector pQE80L and followed with production of mutant HMGB1s and identification by Western blotting.      Results     Six mutant proteins were designed and constructed into prokaryotic expressive vector pQE80L. The recombinant HMGB1 proteins were obtained and identified by Western blotting.      Conclusion     Human HMGB1 mutants have been successfully constructed and the expression and characterization of intent proteins  are identified. It will lay a foundation for further study on biological functions of HMGB1.