Diss. (sammanfattning) Stockholm : Karol. inst. / Härtill 7 uppsatser.

The extant medically and socially important cobras have been the subject to several comparative taxonomic studies since the 1940s, but still lack an inclusive and thorough phylogenetic tree. With recent major advancements in phylogenetic analysis, it is now common to use multiple independent loci for studying the phylogenetic relationships within groups. For the first time, 27 from the 29 identified Naja species, alongside 5 putative new or elevated species had 4426 base pairs across 1701 sequences of mitochondrial and nuclear DNA sequence data analysed. The results continue to support the monophyletic core cobra clade encompassing the genera Walterinnesia, Aspidelaps, Hemachatus, Pseudohaje and Naja (1.0 Bayesian posterior probability (BPP)), in addition to the grouping of four monophyletic subgenera within Naja. The group of African spitting cobras, Afronaja, is positioned as the sister group to the rest of the genus. Moderate support (0.8 BPP) is found for the grouping of the Asian cobras, Naja, with the African non-spitting cobras, Ureaus. The closest relative to the genus Naja is Pseudohaje goldii, a genus and species never before included in phylogenetic analysis, followed by the sister taxa Hemachatus haemachatus. The king cobra continues to be positioned outside the core cobra group, sister to Hemibungarus calligaster. The results support the hypothesis of three independent origins of spitting, once in the monotypic Hemachatus haemachatus, once within the subgenus Afronaja, and the final origin within the Asian cobras, subgenus Naja. The relationships found were broadly consistent with previous studies, with the additional inclusion of more species creating the most comprehensive cobra phylogeny to date. Further molecular analysis, specifically species delimitation, must be undertaken to ascertain the position of the 5 putative new species included in this study.

Snake venoms are a complex mixture of polypeptide and other molecules that adversely affect multiple homeostatic systems within their prey in a highly specific and targeted manner. Amongst the most potently toxic venoms in the world are those of the Australian venomous snakes, which belong almost exclusively to the elapid family. Their venoms posses a number of unique properties by which they target the mammalian cardiovascular and neuromuscular systems and are the focus for the identification of novel pharmacologically interesting compounds which may be of diagnostic or therapeutic benefit. Although much is known about the biochemical properties of Australia snake venoms as a whole, little research attention has focused upon individual components at the molecular level. This thesis describes the cloning, characterisation and comparative analysis of a number of unique toxins from the venom gland of the coastal taipan (Oxyuranus scutellatus) and a total of seven other related Australian snakes. These include the factor X- and factor V-like components of a prothrombin activator that causes a highly coagulable state in mammals. Comparative analysis of the sequences identified in this study, along with recombinant expression of an active form of the factor X-like component, provides important information on the structural, functional and evolutionary relationships of these molecules. Numerous other toxins were similarly identified and characterised including a pseudechetoxin-like protein, multiple phospholipase A2 enzymes and neurotoxin isoforms as well as vasoactive venom natriuretic peptides. Identified transcripts included not only toxin sequences but also other cellular peptides implicated in toxin processing, including a calglandulin-like protein. This thesis is the first description of the majority of these molecules at either the cDNA or protein level, and provides a means to study the activity of individual components from snake venoms and probe their function within the systems they specifically target. This study represents the most detailed and comprehensive description to date of the cloning and characterisation of different genes associated with envenomation from Australian snakes.