Prokayotic Expression and Polyclonal Antibody Preparation of P5 Protein of Rice grassy stunt virus

• 1、Institute of Plant Virology, Fujian Agriculture and Forestry University,fuzhou 350002
• 2、Institute of Plant Virology, Fujian Agriculture and Forestry University,fuzhou 350002

Abstract：Rice grassy stunt virus(RGSV),a member of the tenuivirus, is transmitted by the brown planthopper (BPH), Nilaparvata lugens (Hemiptera, Del-phacidae), in a persistent propagative manner. The function of protein P5 encoded by RGSV is unkown, in this study we amplified p5 gen by RT-PCR, and recombined to prokayotic expression vector pDEST17 by Gateway recombinant technology. The recombinant expression plasmid was transformed into E. coli Rosetta to induce expression of P5-HIS6 fusion protein which was used to immunize the rabbits as an antigen and then the antiserum of P5 was obtained. Testing the antiserum by western blot and ELSA and found that the obtained antiserum was specific and had a better potency. Immunocapture RT-PCR and Dot-blot ELISA meProkayotic Expression and Polyclonal Antibody Preparation of P5 Protein of Rice grassy stunt virusthod to detect the RGSV by using antiserum of P5 was also established. Therefore, the antiserum made in this study provide technical support for the diagnosis of RGSV disease and protein P5 function study.

Keywords： RGSV P5 protein prokaryotic expression pAb preparation