FIG. 2.
Construction of HEV mutants derived from the wild-type genotype 3 swine HEV consensus cDNA clone pSHEV-3. (A) Alignment of nucleotide sequences in the junction regions among the mutants and their group assignments for the in vivo infectivity study in pigs. T7p represents a T7 RNA polymerase core promoter, and A15 represents a 15-nt poly(A) tail. The unique AflII and AvrII restriction sites on pSHEV-3 used for the cloning purpose are also shown. For an explanation of the symbols, see the legend for Fig. 1, panel A. (B) In vitro transcription of full-length capped RNAs from the wild-type infectious cDNA clone pSHEV-3 and five HEV mutant clones. Five microliters of the transcription mixture was separated in a 1% agarose gel to check the quality of the transcription before being intrahepatically inoculated into the liver of pigs. An arrow indicates the in vitro RNA transcripts of expected size.