Preparation and Application of Polyclonal Antibody against Vein Yellows Virus P4 on Chili Pepper Plants

  Objective   Pathogenicity of the pepper vein yellows virus (PeVYV) of genus Polerovirus that caused epidemic infection on chili peppers, Capsicum annuum L., in China was studied to assess the potential threat to the crop, and the polyclonal antibody against the virus prepared.

  Methods   Polyclonal antibody of PeVYV was prepared using the purified recombinant P4 protein. A detection method based on the antibody was established to determine the functions and characteristics of P4. The 471 bp of P4 gene was amplified by RT-PCR using the total RNA of infected chili peppers and cloned into prokaryotic expressing plasmid pDEST17 expressed in E. coli Rosetta by arabinose induction. The recombinant P4 was purified by Ni-NTA chromatography for the antibody preparation and verified by western blotting.

  Results   The recombinant P4 protein was soluble with a molecular weight of approximately 25 kDa. The prepared polyclonal antibody was confirmed by western blot to specifically recognize the protein.

  Conclusions   The polyclonal antibody prepared in this study specifically recognized P4. It could be used to characterize and determine the functions of the protein.