Construction and biological characteristics evaluation of recombinant PRV ex-pressing S protein of PEDV
In order to construct a recombinant pseudorabies virus expressing S protein of porcine epidemic diarrhea virus,the S extracellular region gene of porcine epidemic diarrhea virus epidemic strain was cloned into the transfer plasmid pUC-ABK containing the upstream and downstream homologous arms of gG gene and the resistance screening gene to construct the donor plasmid pUC-ABK-S,and the linear homologous recombination fragment A-S-K-B was obtained.The linear homologous recombinant fragment was electrotransformed into the competent cells containing pPRVBac-GFPTK-gE-gl-11k-28k-to obtain recombinant pPRVBac-S by Red homologous recombination,and finally,the recombinant virus rPRV-S was rescued.PCR and IFA identification results showed that S extracellular region gene was successfully transcribed and expressed,the results of biological characteristics evaluation showed that the recombinant virus could be stably inherited,which pro-vided an experimental basis for the development of porcine epidemic diarrhea virus live vector vac-cine.
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