Abstract
Aiming to use DNA sequence for proper identification of venomous snakes in Saudi Arabia, mitochondrial 12S rRNA gene sequences of Bitis arietans, Cerastes cerastes, C. gasperettii, and Echis coloratus were investigated. Concatenated analysis of 16S and 12S rRNA gene sequences (as mosaic) was also performed to validate the usefulness of using more than one gene marker in molecular taxonomy of venomous snakes. DNA extracted from whole blood samples and 12S rRNA was amplified using PCR. Sequences submitted to GenBank were analyzed for similarity using BLAST and aligned using Clustal X followed by manual editing with BioEdit software. Phylogenetic analysis was generated using PAUP version 4, and the evolutionary distance among different groups was calculated using MEGA5 software for both 12S and 16S rRNA genes. Contrary to the morphological classification, phylogenetic analysis revealed that the collected isolates formed three different statistically supported clusters (B. arietans, C. cerastes, and E. coloratus). Although the combined analysis of 16S and 12S rRNA sequences improved the signal, the C. cerastes and C. gasperettii intermixing have not been resolved. Both single and combined genetic distance analyses showed that Echis is more distant from Cerastes than from Bitis. Further studies are mandatory to evaluate other gene markers in molecular classification.