Isolation of genomic DNA from the nose-horned viper and initial analysis of metalloproteinase genesRubil, Tihomir (Avtor) Pungerčar, Jože (Mentor) Vipera ammodytes ammodyteslong-nosed vipergenomic DNAgene structurenucleotide sequencingsnake venom metalloproteinaseclass III metalloproteinaseevolutionSnake venoms, including that of the nose-horned viper (Vipera ammodytes ammodytes, Vaa), are a valuable source of pharmacologically interesting substances. Vaa venom contains the following major groups of protein toxins: serine proteases, secreted phospholipases A2, snake C-type lectin-like proteins and metalloproteinases. The latter are mainly responsible for hemorrhagic and hemostatic effects of the venom. Of notable reference are metalloproteinases that belong to the P-III (MPIII) class. These metalloproteinases are diverse in both quaternary protein structure as well as in their function. The organization and features of their genes are yet largely unknown. The aim of this thesis was to isolate Vaa genomic DNA, and to amplify and analyze genomic sequences encoding two metalloproteinases, Vaa hemorrhagin 4-A and Vaa metalloproteinase III-like protein 3 (Vaa-MPIII-3). Their presumed nucleotide sequences were amplified by PCR and determined by the Sanger dideoxy-chain termination method. Results indicated that the specific amplification attempts for both genomic sequences were unsuccessful. In addition, a putative contig of 23,133 bp harboring the genomic sequence coding for Vaa-MPIII-3, obtained from the whole Vaa genome sequencing project, was acquired and its structure analyzed. The Vaa-MPIII-3 gene is composed of 10 exons and 9 introns. It has been confirmed that Vaa-MPIII-3 represents a new subtype of metalloproteinases lacking the entire metalloproteinase (enzymatic) domain and the first part of the succeeding disintegrin domain, reflecting a unique evolution of its gene, separated from that of enzymatically active snake venom MPIII.20202020-11-18 07:15:17Magistrsko delo/naloga122031sl