Coexpression for E Gene of Japanese Encephalitis Virus and HA Gene Epitopes of Human Influenza Virus and the Application for the expression Product
Abstract: Using a pair of specific primers designed according to the nucleotide sequence of Japanese encephalitis virus and Hameoaggulatinin of Human influenza virus from GenBank, the gene of JEV E protein’s main antigenic domain including the sequence of Human influenza virus ha gene was amplified with PCR method. The PCR product was successively cloned into pET-32a(+) vector to get a prokaryotic expression plasmid pET-EHA. After the positive plasmid was transformed into the host cell BL21(DE3) , the target gene was successfully expressed in the form of inclusion bodies when induced with IPTG. Western-blotting analysis proved the recombinant protein has good reactivity ability against JEV antibodies. The Latex Agglutination Test (LAT) method for the detection of JEV antibodies was established with the purified recombinant protein. The result indicated that LAT was a simple, rapid, sensitive, specific, inexpensive method which is suitable for detecting antibodies against JEV in serum and serological survey of JE .