FIG. 3.

Construction of the full-length genome plasmid for the chimeric R(G) strain. Two cDNA fragments containing the Nishigahara G-ORF and the RC-HL M-G noncoding region were amplified by RT-PCR and PCR, respectively. The P1 and P12 primers (italics) were designed for a previous study (12). For construction of a PstI site in the G-L noncoding region, site-directed mutagenesis was performed with a RPsdPstI mutagenesis primer. After cloning and sequencing, the two fragments were connected and subcloned into pRC-HL(+).