Expression and purification of rabies virus protein P: production and characterization of anti-P polyclonal antibodies
Keywords:
rabies virus, P protein, protein expression and purification, polyclonal antibodiesAbstract
Rabies virus (RV) phosphoprotein (P) is a multifunctional protein. However, the functions of this protein are
not fully understood. To further investigate this question, the P protein was expressed and used to raise monospecific
polyclonal antibodies. The gene coding for P protein was cloned into expression vector pET-32a (+) to yield pET-32a
(+)-P. His-tagged P protein was expressed in E. coli BL21 cells and analyzed by SDS-PAGE. After purification by nickel
affinity chromatography under native conditions, the recombinant P protein was used to raise anti-P polyclonal
antibodies in mice. Western blot analysis showed that the P protein was recognized by the polyclonal antibodies.
Immunofluorescence assays also showed that the antibody was able to recognize the native P protein in RV-infected
cells.
