Prokaryotic Expression And Purification Of ASFV E183L Gene And Its Polyclonal Antibody Preparation

African Swine Fever(ASF)caused by African Swine Fever Virus(ASFV)has become one of the major diseases that endanger pig breeding industry because it can kill sick pigs quickly.In recent years,it has been introduced into China,which has a great impact on China’s pig industry and pig trading market.However,because the complex structure of ASFV leads to its immune escape mechanism to the host,there is no effective vaccine against ASFV at present,so timely quarantine and elimination is the main way to prevent and control the epidemic spread.At present,it is the primary problem to develop detection technology and materials with strong specificity,high sensitivity and good safety.Efficient and reliable detection means are the key to early successful diagnosis of ASFV and monitoring its spread.Based on the purpose of providing good materials for effective detection,this study analyzed the bioinformatics of ASFV E183 L gene,and made use of an economical and efficient prokaryotic expression system to successfully express the prokaryotic expression vector of p ET-30a-E183 L.After purification,p54 protein was successfully obtained,and polyclonal antibody was successfully prepared by using p54 protein.The results showed that the prokaryotic expression vector p ET-30a-E183 L successfully expressed p54 fusion protein by SDS-PAGE,and the identification results showed that the target band was single,and it was well combined with His tag antibody.The purified p54 protein was used to immunize mice,and the titer of the prepared specific antibody reached1: 6400.P54 protein prepared in this study provides a good raw material for serological detection of ASFV,and provides a reference for the follow-up vaccine development.