Expression and purification of thermolabile hemolysin of vibrio parahaemolyticus

  Objective   To express and obtain the fused protein thermolabile hemolysin(TLH)of vibrio parahaemolyticus (VP).

  Methods   Prokaryotic expression plasmid p ET32a+-tlh was constructed and incorporated into E.coil BL₂₁(λDE₃) the engineered bacteria were inducted by IPTG to express TLH, which was identified by SDS-PAGE and purified by using 6×hisNi-NTA Resin.

  Results   the induced expression fusion protein was inclusion, which expression reached to 10% of the total protein of BL₂₁(λDE₃), the high pure fused protein TLH was obtained by using affinity chromatography method.

  Conclusion   the recombinant plasmid p ET32a+-tlh had stably and efficiently expression in BL₂₁(λDE₃), the high pure fused protein TLH could be obtained by using affinity chromatography method by using low concentration imidazole β,-ME and Triton x-100 to reduce contaminated protein.