Norovirus Narita 104 Virus-Like Particles Expressed in Nicotiana benthamiana Induce Serum and Mucosal Immune Responses
Figure 1
Initial characterization of transient expression of NaVCP. (a) Northern blots of RNA from leaves infiltrated with pICHsNaV (lane 2) showing genomic RNA (gRNA) and subgenomic RNA (sgRNA) of NaVCP after 3 DPI. Lane 1 is noninfiltrated leaf as negative control. Two μg of total RNA was loaded and probed with probe specific to TMV 3′UTR. Ribosomal RNA loading is shown at the bottom. (b) Coomassie stained SDS-PAGE of NaVCP showing presence of NaVCP at expected size ~58 kDa. Lane 1: protein molecular mass markers; lane 2: insect-derived NaVCP (1 μg); lane 3: crude protein extract (15 μg) from leaves infected with empty vector; lane 4: crude protein extract (15 μg) from leaf samples infiltrated with pICHsNaV and harvested at 4 DPI. The 64 kDa and 48 kDa molecular mass markers are indicated by arrows at left. (c) Western blot of SDS-Page of sNaVCP showing presence of NaVCP at expected size ~58 kDa. Lane 1: insect-derived NaVCP (25 ng); lane 2: crude protein extract (10 μg) from leaves infected with empty vector. Lane 3: crude protein extract (10 μg) from leaf samples infiltrated with pICHsNaV and harvested at 4 DPI. (d) Hypersensitive response on pICHsNaV infiltrated region. Part of the leaf that was infiltrated with pICHsNaV displayed cell death 5 DPI whereas such symptoms are absent in infiltrated areas of GFP, NVCP, and empty vector (Neg). Inset shows GFP fluorescence at 10 DPI.
