Prokaryotic Expression of Porcine Mydoid Differentiation Factor 88

Porcine MyD88 molecule fragment(pMyD88-ED)was amplied by PCR method from the pGEM-T-pMyD88 recombinant plasmid,the pMyD88-ED was cloned into pET-30a vector and expressed in Escherichia coli.The recombinant strain was induced in different IPTG concentrations and different time.The results showed that the optimum concentrations of IPTG is 0.05 mmol/L,the optimum time is 6 h.SDS-PAGE analysis showed that the recombinant protein was expressed with a molecular weight of 35 ku and existed mainly in the form of inclusion bodies.The protein was purified by elution methods and the purity reached 90%.The Western-blot analysis indicated that the hybridization bands is appeared around 35 ku,and it showed that the fusion protein was identified by the monoclonal antibody of HIS label protein,so the target protein was expressed successfully in the Escherichia coli.