Figure 1.

Rotavirus (RV) genes expressed from herpes simplex virus type 1 (HSV-1) amplicon vectors. (a) Schematic representation of the HSV-1 amplicon vectors expressing or co-expressing RV genes VP2, VP6, and VP7. Polycistronic expression is facilitated by two picornavirus internal ribosome entry site (IRES) and an encephalomyocarditis virus (EMCV)-derived IRES, and controlled by the HSV-1 IE4/5 promoter. All vectors contain the enhanced green fluorescent protein (EGFP) reporter gene to support titration of vector stocks. The HSV-1 origin of DNA replication (ori_S) and packaging/cleavage signal (pac) are indicated. The polyadenylation signal was from SV40. (b) Vero 2-2 cells were infected with the indicated HSV-1 amplicon vectors (MOI 1), and total cell lysates were harvested at 24 hours postinfection (hpi). Transgene expression was analyzed by western blot using a polyclonal rabbit anti-RV serum for detection of RV proteins or a monoclonal anti-GFP antibody to stain EGFP. Detection of actin was used as loading control. Purified and inactivated wt RV strain SA11 served as positive control. The positions of molecular weight markers (kDa) are indicated.