Molecular cloning and physical mapping of bovine herpesvirus 4 strain DN 599 and comparison with two American field-isolates

Ninety four percent of the genome of bovine herpesvirus 4 (BHV-4) strain DN 599 was cloned and a physical map was constructed by Southern blot analysis using a library of cloned fragments cleaved with the 3 restriction enzymes (Eco RI, Bam HI, and Hin dIII). The genome length was estimated to be 156.5 kbp +/- 0.7. The genome comprises a region of unique segment (114 kbp) and two flanking segments containing tandem repeats. The size of each repeat was approximately 2.35 kbp and each repeat contained one Eco RI site and two Bam HI sites. We also examined two recent American field-isolates of BHV-4 and compared the Eco RI maps of the two isolates with that of DN 599. We observed the following: (1) insertions or deletions of restriction sites at the periphery of the unique segment; (2) variation in the lengths of junction fragments; (3) variations in the lengths of hypermolar Eco RI fragments containing the repeats; and (4) the Eco RI map of one of the American field-isolates resembles the BHV-4 "Movar type" of Europe.