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DEVELOPMENT OF DNA BASED MICROSATELLITE MARKER TECHNOLOGY FOR STUDIES OF GENETIC DIVERSITY IN CENTRAL STONEROLLER (CAMPOSTOMA ANOMALUM) POPULATIONS
Citation:
Dimsoski, P. AND G P. Toth. DEVELOPMENT OF DNA BASED MICROSATELLITE MARKER TECHNOLOGY FOR STUDIES OF GENETIC DIVERSITY IN CENTRAL STONEROLLER (CAMPOSTOMA ANOMALUM) POPULATIONS. Presented at Society of Environmental Toxicology and Chemistry, Philadelphia, PA, November 12-14, 1999.
Description:
The level of genetic diversity of aquatic species is a critical indicator of stream system condition for which few data exist. There is strong evidence suggesting that environmental stressors affect the genetic diversity of exposed populations. In order to study genetic diversity, we have developed a set of DNA microsatellite markers. The markers are dinucleotide (CA) repeats which are assumed to be randomly distributed across the genome of Campostoma anomalum (Central Stoneroller). A high level of polymorphism, which is essential for genetic diversity and monitoring studies of populations, is generated by the variable number of nucleotide repeats. A microsatellite-enriched genomic library was constructed, followed by isolation of CA-repeat DNA sequences that were cloned into E. coli. Over 50% of the colonies were 'positive' for microsatellite inserts. The unique flanking regions and dinucleotide repeats of insert-containing colonies were sequenced from both ends of the insert. All of the inserts previously identified as 'positives' contained CA-repeats. The number of repeat units on the 22 sequenced microsatellites ranged from 10 to over 100. It is expected that a higher level of polymorphism will be associated with the higher number of dinucleotide repeats. A set of unique microsatellite primers was designed and prepared for automated genotyping and sequencing. Ten of the 22 designed primers yielded PCR products of the expected size. The markers will be used to characterize the level of heterozygosity for Central Stonerollers across a range of stressor exposures in regional studies of stream ecosystem condition in the Eastern Untied States.