Prokaryotic Expression of African Swine Fever Virus pE248R Protein and Preparation of Polyclonal Antibodies
In this study,the 747 bp E248R gene was amplified by PCR from inactivated samples of African swine fever virus(ASFV),and the prokaryotic expression plasmid pCold Ⅰ-pE248R was constructed by homologous recombination method.After induced with 1 mmol/L IPTG for 16 h,the recombinant protein was identified with a molecular mass of 32 kDa in SDS-PAGE.The purified pE248R recombinant protein was used to immunize mice 4 times to prepare anti-pE248R polyclonal antibodies.Then the polyclonal antibodies were used to detect the recombinant viruses rPRRSV-E248R,which was constructed and rescued by our laboratory and had been proved to stably express ASFV pE248R protein.The results showed that the prepared polyclonal antibodies specifically bound to rPRRSV-E248R.The availability of the polyclonal antibodies against the prokaryotic pE248R protein laid a foundation for developing a rapid and specific serological detection method for ASFV E248R gene,and also provided a research foundation for the functional study of pE248R protein.
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