Expression and purification of rabies virus protein P: production and characterization of anti-P polyclonal antibodies

Authors

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.

DOI

10.56808/2985-1130.2909

First Page

257

Last Page

264

Recommended Citation

Pan, Yan; Meng, Xian-Ming; Wei, Xian-Kai; Li, Jun; Lu, Zhuan-Ling; Tang, Hai-Bo; Liang, Jing-Jing; and Luo, Ting Rong (2018) "Expression and purification of rabies virus protein P: production and characterization of anti-P polyclonal antibodies," The Thai Journal of Veterinary Medicine: Vol. 48: Iss. 2, Article 14.
DOI: https://doi.org/10.56808/2985-1130.2909
Available at: https://digital.car.chula.ac.th/tjvm/vol48/iss2/14