Objective: To clone and construct expression vector pET32a+-tlh to acquire biologically functional thermolabile hemolysin of Vibrio parahaemolyticus.
Methods: tlh gene was cloned and the expression vector pET32a+-tlh constructed. The tlh gene of Vibrio parahaemolyticus was expressed in DE3 in the form of inclusion body, which was resolved in 8 mol/L urea followed by purification of the fusion protein using affinity chromatography and renaturation through gradient dialysis, protein concentration reduction and oxidoreduction.
Results and conclusion: The purified and renatured protein possessed hemolytic and immunogenic activities.