Sequencing the control region of the male-transmitted mitochondrial genome of the horse mussel, Modiolus modiolus
LE3 .A278 2013
Bachelor of Science
Marine mussels (Family Mytilidae) have a unique system of mitochondrial DNA (mtDNA) inheritance, called doubly uniparental inheritance (DUI). In this system, two mitochondrial genomes exist: the F genome, that is female-transmitted, and the M genome, that is male-transmitted. It has been proposed that the mitochondrial control region of marine mussels could play a role in sex determination. In most animals, the control region plays an important part in regulation of transcription and replication of mitochondrial genes, but is not known to perform any other function. My objective was to develop a procedure to amplify and sequence the control region from the male genome of Modiolus modiolus, commonly known as the horse mussel. Initially, mtDNA from M. modiolus sperm was isolated, and the cytochrome c oxidase subunit I (COI) and 16S ribosomal RNA genes were amplified using PCR. Amplification was verified through agarose gel electrophoresis, and samples were sent to McGill University for sequencing. These genes are thought to flank the M. modiolus control region and surrounding genes, based on gene order in female M. modiolus. Partial COI and 16S rRNA sequences were used to design primers for the male control region and surrounding mtDNA using the Primer3 program. Nine primers were designed to target both sides of the genome between COI and 16S rRNA. Using long PCR, three different pairs of primers amplified a region of approximately 10 kb, which is close to the size of the known female control region and surrounding genes. Further sequencing will be required to determine if any of the samples contain the male control region. Analysis of the male control region sequence will help provide valuable information toward further exploration of its function and the evolutionary dynamics of this gene within the family Mytilidae.
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