Packaging of Rift Valley fever virus pseudoviruses and establishment of a neutralization assay method

Li, Yuetao; Zhao, Yongkun; Wang, Cuiling; Zheng, Xuexing; Wang, Hualei; Gai, Weiwei; Jin, Hongli; Yan, Feihu; Qiu, Boning; Gao, Yuwei; Li, Nan; Yang, Songtao; Xia, Xianzhu

To confirm that the acquired viral particles were RVFV pseudoviruses, western blotting was performed to determine whether the pseudovirus envelope protein was composed of RVFV structural proteins. The supernatants of cells transfected with pcDNA3.1-M-rvfv were examined by using a rabbit polyclonal anti-RVFV-MP12 antibody (Fig. 4) . The western blotting results showed two specific bands, with molecular weights of approximately 65 kDa and 56 kDa, consistent with the sizes of the RVFV G1 and G2 proteins, respectively. The negative control group did not show specific bands. These results indicated that the target protein was correctly expressed. Moreover, the western blotting detection results both indicated that the obtained RVFV pseudoviruses contained the RVFV G1 and G2 proteins and confirmed that pseudoviruses were successfully established.