Yan Pan, Xian-Ming Meng, Xian-Kai Wei, Jun Li, Zhuan-Ling Lu,Hai-Bo Tang, Jing-Jing Liang, Ting Rong Luo*

Abstract  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.
Keywords: rabies virus, P protein, protein expression and purification, polyclonal antibodies