关键词: 干扰素&beta, 原核表达, 包涵体

Abstract:

Objective To establish the prokaryotic expression model of IFN-β by constructing the prokaryotic expression vector of recombinant mouse IFN-β and screening the conditions for its inclusion body solubilization,and to provide experimental basis for screening the proper solubilization conditions.Methods DNA was extracted from fresh liver tissue of mice.The prokaryotic expression vector of mouse IFN-β was constructed by traditional methods and was named IFN-β′.Based on the traditional method of solution,the inclusion body dissolving process conditions were improved,such as solution composition pH environment,bacteria and dissolved liquid ratio,and the effect of dissolved temperature  and the dissolution time of inclusion body.Results The mature IFN-β′protein was identical with the predicted results and the induced expression was normal.The changes in pH and DTT  in dissolved liquid had no effects  on inclusion body solubilization.The inclusion body could not be dissolved by washing liquid after liquid washing.Temperature had almost no effect on inclusion body solubilization.When the ratio of dry bacteria to solution was increased from 1 g∶5 mL to 1 g∶20 mL,the IFN-β′protein inc
lusion body could be partially dissolved.Conclusion The recombinant mouse IFN-β prokaryotic expression vector is constructed successfully.The target protein is expressed with the form of inclusion body but not easy to be dissolved,and the  dissolved liquid proportion is the only condition to affect the dissolution of in clusion body protein among the screened inclusion body solubilization conditions.
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Key words: interferon-β, prokaryotic expression, inclusion body
ion, inclusion body