Isolation And Identification And Non-coding Short Segments Of DNA Library Construction Of Swine Pseudorabies Virus Strain

A strain of pseudorabies virus (PRV) was isolated from tissues of dead pigletswith PRV-like diseases from a farm in Linzhou Henan. The isolate was able topropagated in cells of PK-15and caused to typical cytopathogenic effect aftercontinuous cell culture and be neutralized by PRV positive sera in microneutralizationtest. The tissue culture infective dose (TCID50) was10-5.3/0.1mL from thedetermination of the half cells infected with the virus. The isolated PRV wassusceptible to chloroform and ether,and could be inactivated in56℃water bath for30min.The pseudorabies clinical symptoms were displayed on rabbits and miceinoculated with the isolated virus after a few days. According to PRV gp50geneconserved sequence of GeneBank the primers were designed.The gene fragmentsamplified by polymerase chain reaction (PCR) were recycled, purified and sequencedby sequencing company. Sequence comparison analysis with other classic PRV strainsby using DNA star software, the homology of the isolate and other classic PRV strainswas99%. The biological characteristics of the isolated strain be further analysis andresearch.According to GanBank (sequence number: JF797219) typical PRV straincomplete genome sequence designed multiple pairs of specific primers using PrimerPremier5software in the non-coding region. Through the optimization of PCRamplification method in non-coding region of short segments of DNA, the amplifiedproducts were purified and cloned to vector and successfully constructed a non-codingregion of DNA library. In this study, the isolation and identification of swinepseudorabies virus and non-coding DNA library construction provided importantresearch materials for the gene mutation, virulent variation, virus latent infection andregulatory sequences of the non-coding region.