Construction and Application of Recombinant Herpesvirus of Turkey Expressing HA Gene of Subtype H9 Avian Influenza Virus
In order to construct the recombinant herpesvirus of turkey(HVT)expressing HA gene of H9 subtype avian influenza virus(AIV),based on a Fosmid library continuously covering the whole genome of HVT,the optimized HA gene carrying a Pec complex promoter was inserted between the loci of HVT053 and HVT054(95 318～95 319 nt)in HVT non-essential region for replication using Red/ET homologous recombination technology and Gateway LR cloning system,a recombinant HVT strain(rHVT-H9-HA)expressing HA gene was obtained and characterized.The results showed that the entry plasmid pEntr-HA containing HA gene was successfully constructed.pFosC HA was formed by transferring the expression cassette containing HA gene between No.53 and No.54 gene of the pFosC DEST cosmid by LR reaction;after extracting 5 cosmids,the rescued recombinant virus rHVT-H9-HA strain was obtained by transfecting secondary-generation CEF cells.Consecutive cell passages and animal tests showed that the rHVT-H9-HA strain was with high virus titer,genetic stability and anti-interference against maternal antibodies,and capable of fighting against H9 subtype AIV with long immunization duration;compared to inactivated vaccines,the effect of"one injection against two diseases"was better.A technical support was provided by the study to develop the recombinant HVT live-vector vaccine for H9 subtype AIV.
recombinant rHVT-H9-HA strainHVTHA geneRed/ET homologous recombination technologyGateway LR cloning system
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