Complete Analysis Of JEV/HW Strain And The Construction Of Its Gene Engineering Vaccines

Based on the HW strain of Japanese Encephalitis Virus (JEV) isolated by Hubei Academy of Agricultural Sciences on Wuhan region of Hubei Province, We determined the complete genome of the JEV/HW strain and its amino acid sequence was deduced, Accession number of HW strain is AY849939. The complete nucleotide sequence is composed of 5' nontranslated regions(5'NTRs), C, PrM, E, NS1, NS2a, NS2b, NS3, NS4a, NS4b, NS5 and 3 NTRs. Phylogenetic tree was constructed based on 18 JEV strains.First, the complete genome including eight fragments of JEV/HW Strain were amplified by RT-PCR. Comparison complete nucleotide sequence of HW strains with other 18 typical JEV isolates revealed high level sequence identity. Ranging from 88.7% (KV1899) to 99.3%(P3) for nucleotide sequence and ranging from 97.0% (KV1899) to 99.4%(P3) for protein sequence. Our results showed that JEV/HW strains was derivative virus of JEV/P3 strain. It maked the base of prevention of JEV in the Hubei province.Recombinant Prokaryotic expression plasmids of pGEX-KG-E and pGEX-KG-NSl were constructed, DNA Vaccine Vectors of pcDNA3.1-E and pcDNA3.1-NSl were also constructed, Prokaryotic expression plasmids of pGEX-KG-NSl was expressed on E.coil BL21 (DE3).Additionally, Study of JEV/HW strain reproduction with cell and mice was developed. These studies laid the base of prevention of JEV in the Hubei province.