부산대학교 도서관

This thesis describes the design, development and adaptation of molecular techniques for the study of mitochondrial DNA (mtDNA). The techniques were used to further understand the levels and evolution of sex-specific mtDNA (F and M genomes) in Mytilus spp. and the presence and distribution of post-deletion mtDNA in the oncocytes of a benign human Salivary Gland tumour (Warthins tumour). The development of two sets of same-site, genome-specific PCR primers allowed for optimisation of a rapid approach for testing neutral evolution of the Mytilus F and M genomes. For both Mytilus edulis genomes, RFLP and SSCP data were used in as input for neutrality tests. The results of these tests agreed with previous findings that the M genome has evolved in a nearly neutral fashion but the F genome is under a higher constraint. These conclusions and the function of the M type COX 3 subunit are further discussed after analysis of F and M COIII nucleotide sequence and the theoretical tertiary structure and positions of replacement amino acids. One of the primer sets was further used in the development and application of a quantitative PCR (Q-PCR), approach to estimate the level of the F and M genomes in somatic tissue, gonad and sperm of Mytilus edulis males. The results allowed for formation of a hypothesis to explain the stochastic mechanism leading to sex determination in Mytilus. Q-PCR was also used to determine the presence and level of a 4977 bp mtDNA deletion in smoker and non-smoker normal elderly human Parotid gland tissue and oncocytes of Warthins tumour, for which the level was significantly increased.