UBC Theses and Dissertations
Japanese quail microsatellite loci amplified by chicken-specific primer pairs Pang, Shirley Wing Yee
Japanese quail (Coturnix japonica) is an important food species as well as a popular animal model for research. Yet, despite their importance, very little is known about their genome. Only allozyme markers have been used to determine the genetic variation of Japanese quail populations and the extent of the protein polymorphism seemed to be relatively low. The use of more variable DNA markers would provide a better understanding of the population structure of these birds. Recently, some scholars suggested that microsatellite markers could detect much higher variation in populations where the detectable allozyme variability is low. Microsatellite loci are often conserved among related species and successful cross-species amplification using heterologous microsatellite primers have been shown in many closely related species of mammals and birds. Microsatellite markers have not been developed for the Japanese quail. The purpose of this study was to examine whether polymorphic microsatellite primers from chicken could amplify corresponding microsatellite loci in Japanese quail DNA templates. Forty-eight chicken microsatellite primer-pairs were tested on Japanese quail genomic DNA using polymerase chain reaction (PCR). Post-PCR DNA fragments were first analyzed on 2% agarose gels and then on Spreadex™ gels. Amplification products were obtained from 28 chicken microsatellite primer-pairs (58.3%) after optimizing the PCR condition for each primer set. Of the 28 primer-pairs that gave PCR products, 12 (25.0%) generated specific products and 16 yielded non-specific amplification products. Specific amplification products from each of the 12 PCR primers were sequenced. Nine of these markers were polymorphic and 3 were monomorphic from 4 Japanese quail populations. Seven of the 9 polymorphic and one of the 3 monomorphic markers contained microsatellite repeats. Three microsatellite loci were confirmed to be homologous to their corresponding loci in chicken. The results indicate the possible use of chicken-specific microsatellite primers in the analysis of Japanese quail genome. Furthermore, results from cross-species characterizations are also presented in the hope of stimulating interest in the use of these markers in various genetic studies.
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