FIG. 8.
Identification of marker mutations in recombinant virus. Various icMHV-A59#1 or wild-type virus amplicons were subcloned into Topo II vectors and sequenced. (A) icMHV-A59#1 sequence across the mutated 3512 Esp3I site; (B) icMHV-A59#1 sequence across the mutated 48 Esp3I site and the MHV A/B junction; (C) icMHV-A59#1 sequence across the mutated 17 Esp3I site; (D) icMHV-A59#1 sequence across the RsrII site and MHV F/G junction; (E) wild-type MHV-A59 sequence across the same domain as shown in panel D. Overlined sequences include the mutated restriction sites and the corresponding wild-type nucleotides at these positions. In panels B and D, the bracketed sequences represent the MHV A/B and F/G junction domains, respectively. Note that all evidence of the Esp3I site, which had been engineered into the sequence, is gone.