LE3 .A278 2015

2015

Stewart, Don

Acadia University

Bachelor of Science

Honours

Biology

Biology

There exists an unusual system of mitochondrial DNA (mtDNA) transmission in many bivalves known as doubly uniparental inheritance. For example, in the blue mussel, Mytilus edulis, mtDNA transmission is not exclusively maternal; male mussels possess both maternally and paternally-transmitted mtDNA genomes. Fathers produce sperm containing mostly male-transmitted (M-type) mtDNA, which they pass through their sons. Mothers produce eggs containing mostly female-transmitted (F-type) mtDNA, which they pass on through their daughters. While the M-type mtDNA is only expressed in the germ line of the male mussel, the F-type mtDNA is expressed in the germ line of the female mussel, and the somatic tissue of both male and female mussels. Occasionally, an F-type mtDNA and a standard M-type (SM) mtDNA genome recombine and the resulting chimeric genome “invades” the paternal route of mtDNA inheritance, giving rise to a so-called “recently masculinized” (RM) mitotype. Quantitative polymerase chain reaction (qPCR) is a technique used to quantify nucleic acids in a sample of interest. This technique differs from conventional PCR in that it monitors the process of amplification in real time. The amplification curves obtained from the process can be used to calculate the initial concentration of template. Due to the high sensitivity of the qPCR, the primers used should only amplify the desired gene while avoiding primer dimer formation. The purpose of this study was to design qPCR primers that would selectively amplify the SM, RM, and F mitotypes of M. edulis. Novel qPCR primers were designed, and a qPCR protocol was optimized. The development of qPCR primers will facilitate future research into the reproductive biology and fitness of alternative mitochondrial DNA polymorphisms in blue mussels.

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https://scholar.acadiau.ca/islandora/object/theses:1245