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. 2019 Nov 14;13(11):e0007800. doi: 10.1371/journal.pntd.0007800

Fig 3. Construction and characterization of a recombinant RABV-based vector displaying virus membrane-anchored BAFF (RABV-ED51-mBAFF).

Fig 3

(A) RABV is a molecular clone of the attenuated vaccine strain of RABV, SAD-B19. The gene encoding ED51-mBAFF from pcDNA-ED51-mBAFF (Fig 1) was digested with BsiWI and NheI and then ligated into pRABV previously digested with BsiWI and NheI, resulting in pRABV-ED51-mBAFF. Infectious virus was recovered and named RABV-ED51-mBAFF. (B) To analyze incorporation of ED51-mBAFF into RABV-ED51-mBAFF virions, sucrose-purified RABV or RABV-ED51-mBAFF particles were separated by SDS-PAGE and transferred to a nitrocellulose membrane. Blots were probed with either an anti-BAFF antibody (left panel) or anti-RABV G antibody (right panel). (C) BSR cells were infected with RABV or RABV-ED51-mBAFF at a MOI of 5 (one-step growth curve, right panel) or 0.01 (multi-cycle growth curve, left panel). Aliquots of tissue culture supernatants were collected at various times post-infection and viral titers were determined in duplicate.