Abstract Peste des petits ruminants (PPR) is an acute infectious disease caused by Peste des petits ruminants virus (PPRV), which seriously endangers the healthy development of the world's sheep industry. Since 2013, PPR has been epidemic in many places in China, and the sheep industry suffered huge losses. In this study, the specific primers were designed according to the complete sequence of N gene of PPRV Nigeria 75/1 strain (GenBank No. HQ197753) in GenBank, and the full-length sequence of N gene of PPRV Wuwei strain (GenBank No. KY429031) was amplified. Based on sequencing and sequence analysis, the prokaryotic expression vector pET-PPRV-N was constructed and then was transformed into Escherichia coli Rosetta (DE3). Subsequently, the recombinant protein His-PPRV-N was purified and the Balb/C mice (Mus musculus) were immunized. Then the anti-serum against N protein of PPRV was prepared. Finally, the immunogenicity of recombinant protein was analysed by ELISA , Western blot and IFA (immunological fluorescence assay). The results showed that the N gene of PPRV Wuwei strain shared 99.0% homology with 24 PPRV strains isolated from China as well as Mongolia strain 2016. The phylogenetic tree analysis identified that the PPRV Wuwei strain belonged to lineage Ⅳ. The result of SDS-PAGE showed that the recombinant protein His-PPRV-N was successfully expressed in E. coli Rosetta (DE3) with the form of soluble protein, and the relative molecular weight was approximately 57.8 kD. The results of ELISA ,Western blot and IFA indicated that the His-PPRV-N possessed excellent immunogenicity. The results provide basic information for the establishment of the detection method of PPRV and for further study on biological functions of PPRV N protein.
WANG Wen,BAO Shi-Jun,XING Xiao-Yong, et al. Prokaryotic Expression and Immunogenicity Analysis of N Gene of PPRV Wuwei Strain Isolates[J]. 农业生物技术学报, 2019, 27(8): 1478-1484.
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