Preparation and Application of Polyclonal Antibodies Against Nucleocapsid Protein N and Envelope Membrane Glycoprotein Gn of Tomato Spotted Wilt Orthotospovirus

Tomato spotted wilt disease caused by tomato spotted wilt orthotospovirus (TSWV) has resulted seriously yield and quality losses in vegetables and flowers in China. In this study, nucleocapsid protein N gene and membrane polysaccharide protein Gn gene of TSWV were cloned into prokaryotic expression vectors, respectively. The plasmids were transformed into Rosetta(DE3) cells for expression of N or Gn proteins, which were used as immunogens to immunize New Zealand white rabbits, then the polyclonal antibodies against N and Gn proteins were successfully prepared. Western blot analysis showed that the polyclonal antibody against N protein could bind to the N protein specifically. Indirect ELISA showed that the titer of antiserum against N protein was about 1:12 800. However, the specificity of antibody against Gn protein was poor, and the titer was not detected. The polyclonal antibody against N protein has been prepared successfully, and it can be a useful reagent for disease diagnosis in field, the research of virus localization in insect body and the interaction of virus and insect vector.