Cloning and molecular characterization of the myxoma virus genome

Restriction enzyme cleavage maps of the genomes of the Uriarra (Ur), Glenfield (GV), and Lausanne (Lu) strains of myxoma virus were deduced for the enzymes EcoRI, KpnI, BamHI, SalI, HindIII, BglI, PstI, and PvuII. Restriction maps for the three strains were indistinguishable with the exception of an additional KpnI site in the Lu genome at map position 38.8. Genomic DNA fragments were cloned into the plasmid vector pGEM-3 and the viral genome was determined to be 163.6 (+/- 0.2) kb in length. Covalently closed terminal fragments were identified by electrophoresis of "snapback" fragments and the 5.3 kb BglI end fragment was cloned after S1 nuclease digestion of the hairpin structure.