Expression Of ASF-VP72 And Establishment Of ELISA Mediated Detecting Method To African Swine Fever

African swine fever (ASF) is a highly lethal viral hemorrhagic disease in domestic pigs. The morbidity and mortality of the disease caused by virulent virus can be 100%, with characteristic of multiple hemorrhages, breathing difficulty and nerve symptoms. It presents a great potential harm to swine industry worldwide. China is presently free of the disease, but the risk of its entrance is possible due to increasing international trade. Thus, it is necessary to develop quick and accurate diagnostic methods in advance.The VP72 is the major and conserved structural protein of ASFV, which is commonly used as the antigen for diagnostic purpose. To establish an ASF-specific antibody detection methed, the VP72 gene and its soluble form (VP72S) were amplified from the plasmid containing the gene. After digestion by EcoRI and SalI, the PCR products were inserted to pUC20 vector for sequence analysis, which showed that the amplified VP72 gene was identical to that of ASF BA71 strain. The pUC-VP72(S) was digested by restriction enzymes and then subcloned into the prokaryotic expression vector pGEX-6p-1. The GST-VP72(S) was expressed in E.coli by IPTG induction, which was confirmed by western blotting .The anti-serum was prepared by injecting mice with eukaryotic expression vector pcDNA-VP72. An indirect ELISA was...