The molecular phylogeny of macropodid herpesviruses 1 and 2 (MaHV-1 and -2) has been investigated by cloning and sequencing the genes encoding glycoprotein B from both viruses. Phylogenetic reconstructions based on the putative amino acid sequences of glycoprotein B indicate that MaHV-1 and -2 are most closely related to the subfamily Alphaherpesvirinae. Within the Alphaherpesvirinae, MaHV-1 and -2 are closely associated with those herpesviruses that infect primates. This phylogenetic relationship does not fit the constraints of the proposed co-evolution theory described for other members of the Alphaherpesvirinae which have mammalian hosts.Macropodid herpesviruses (MaHVs) have been implicated in fatal disease outbreaks amongst the captive marsupial populations of Australia (Finnie et al., 1976 ;Dickson et al., 1980). These outbreaks have resulted in the isolation of nine MaHVs which have been classified into two species called macropodid herpesvirus 1 and 2 (MaHV-1 and MaHV-2) (Johnson & Whalley, 1990). Serological evidence indicates that these viruses are widespread among Australian kangaroos and wallabies (family Macropodidae) (Webber & Whalley, 1978 ;Wilks et al., 1981). MaHV-1 and -2 are represented by isolates from the parma wallaby (Macropus parma) and the dorcopsis wallaby (Dorcopsis meulleri luctuosa), respectively (Finnie et al., 1976 ;Wilks et al., 1981).The family Herpesviridae is a large group of viruses that have double-stranded DNA genomes. Biological characteristics such as host symptoms, site of replication and site of e-mail mahonyt!dpi.qld.gov.auThe nucleotide sequence data for glycoprotein B from MaHV-1 and -2 reported in this paper have been deposited in GenBank and assigned the accession nos AF061754 and AF061755 respectively. latency have been used to describe three major subfamilies, Alpha-, Beta-and Gammaherpesvirinae, within the family Herpesviridae. Biological characteristics have been used to place MaHV-1 and -2 within the subfamily Alphaherpesvirinae.The Herpesviridae is the best characterized at the molecular level of all the large DNA virus families. As a result the molecular phylogeny of the Herpesviridae has been well studied (McGeoch et al., 1995). Current models of mammalian Alphaherpesvirinae evolution indicate that members of this subfamily have co-evolved with their respective hosts (McGeoch et al., 1995). Consequently, this hypothesis predicts that the molecular evolutionary pattern of the Alphaherpesvirinae reflects that of their hosts. Importantly, the molecular datum used to elucidate these relationships has been derived from Alphaherpesvirinae which infect eutherian mammals. Absent from this literature is the molecular phylogenetic position of alphaherpesviruses which infect metatherian mammals (marsupials). It has been estimated that eutherian mammals and marsupials diverged from a common ancestor approximately 130 million years before present (MYBP) (Janke et al., 1997) whereas the divergence of the Alphaherpesvirinae which infect eutherian mammals from a common ances...