Isolation And Identification Of Avian Encephalomyelitis Virus Henan Strain And Cloning And Sequence Analysis Of The VP0 Gene Of The Strain

One virus was isolated from brain of infected chicken with conspicuous neural symptoms by continuously and rapidly passaging in special-Pathogen-Free (SPF) chicken embryos.the virus was resistant to acid ,chloroform, trypsin and heat(50 癈)under the protection of two- valent magnesium ion.The virus titer was 10~5 /0.2ml and neutralization index was 72.44.A pair of primers for avian encephalomyelitis virus (AEV) isolate HN was designed according to published AEV genome. With use of reverse transcription-Polymerase Chain Reaction(RT-PCR),fragment VPO was in vitro amplified from SPF chicken embryo tissue after inoculation of AEV isolate HN. The fragment VPO gene was inserted into plasmid PGEM-T-Easy to obtain the recombinant plasmid VPO-PGEM-T.VPO gene sequence was 242aa. By comparison with other published gene of AEV, the nucleotide sequence and deduced amino acid of VPO of the strain were and homoloyies,with the correspondent sequences of AEV strain BZ and HB. Respectively ,This demonstrated that the VP)Gene was conservative. The experiment offered theoretical foundation to the studying of structure and function of AEV VPO gene,molecular epidemy and engineered vaccine.