Prokaryotic expression of lumpy skin disease virus P32 protein and preparation of polyclonal antibodies
To express bovine Lumpy Skin Disease Virus(LSDV)P32 protein and the preparation of its polyclonal antibodies,the LSDV-P32 gene sequence in GenBank was downloaded,followed by bioinformatics analysis of the physicochemical and biological properties of the P32 protein and ligation of the B-cell epitope-rich region to the pCold-TF vector.After optimizing the expression conditions,purified recombinant protein was identified by SDS-PAGE and Western blot.Purified protein was used to immunize rex rabbits to prepare polyclonal antibodies against P32 protein,and properties of polyantibodies were identified by ELISA,indirect immunofluorescence assay(IFA)and Western blot.The results showed that the soluble LSDV-P32 protein was successfully expressed,and the optimal expression conditions were 120 r/min shaking for 30 h at 15℃with 0.1 mmol/L IPTG.The rabbit-derived polyclonal antibodies with a titer of 1∶409 600 in ELISA,and can be used for ELISA,Western blot,and IFA.This study provides technical support for the functional study of LSDV-P32 protein and the development of related detection reagents in future.
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