Isolation And Identification Of Orf Virus Anhui Strain And Prokaryotic Expression Of Its B2L Gene

Orf virus is also called contagious ecthyma,which is an highly addicted epithelial and highly contagious disease of sheep and goat.Orf virus is included in the Para-poxvirus genus of the Poxvindae family.Orf virus distributed all over the world,which captured people' attention recent years.Attenuated vaccine and inactivated vaccine is used most widely to prevent from this disease.But vir?Lence return and the emergence of virus variants insecurity were often happened.So it is necessary to find some measures to prevented this disease.Orf virus was isolated from the suspicious scab materials of lambs which was collected from Anhui.Hela cell was used to isolate Orf virus.On the basis of the published nucleotide sequence of ORFV B2 L gene.The B2 L gene's specific pair of primers were designed.The B2 L gene was amplified by PCR,then the phylogenetic tree is constructed.The B2 L gene was about 1137 bp long,it shared 99% similarities with the reference sequence which is submitted to NCBI GenBank,and it had high homology with the Liaoning strain(HQ694773).Then the B2 L gene was cloned into pET-32 a vector,pET-32a-B2 L recombinant prokaryotic expression vector was constructed,and was successf?Lly expressed in E.coli.BL21,The product was detected by SDS-PAGE and Western blot.Purified recombinant B2 L protein was used as an antigen for BALB/C mice to make polyclonal antibody.