Distribution of restriction enzyme sites on the bovine parvovirus genome and comparison to other autonomous parvoviruses

A physical map has been constructed for bovine parvovirus (BPV) replicative form (RF) DNA synthesized in vitro. Analysis of restriction enzyme digestion products for 12 enzymes by neutral gel electrophoresis allowed the mapping of 26 cleavage sites between the 3 ' and 5 " termini of the in vitro RF DNA. Cleavage sites for these nzymes were not detected within the T-terminal hairpin. HindlII, SalI, SmaI, SstlI and XorlI did not cleave BPV DNA, while 12 other enzymes cleaved the genome at one to four sites. Eleven additional enzymes were shown to cleave the genome at 8 to 20 sites. Comparison of the restriction site locations,for BPV with those of other autonomous parvoviruses suggests that there may be conservation within the genome of certain restriction sites. Bovine parvovirus (BPV) is an autonomous parvovirus with a linear, single-stranded DNA genome of approximately 5550 bases (Snyder et al., 1982). The 3'-terminal duplex of parvovirus DNA has been shown to function in vitro as a template-primer for prokaryotic DNA polymerases, allowing conversion in vitro of single-stranded viral DNA to a linear, double-stranded replicative form (RF) (Bourguignon et al., 1976; Salzman et al., 1978; Banerjee t al.