Aquaculture Genetics and Breeding Laboratory, Ohio State University Aquaculture Research and Development Integration Program, 1864 Shyville Road, Piketon, Ohio 45661, USA.
Department of Chemical & Biomolecular Engineering, Ohio State University, 140 West 19th Ave., Columbus, Ohio 43210, USA.
Int J Mol Sci. 2009 Jan;10(1):18-27. doi: 10.3390/ijms10010018. Epub 2008 Dec 27.
To perform whole genome scanning for complex trait analysis, we isolated and characterized a total of 21 novel genomic-SSRs and EST-SSRs for yellow perch (Perca flavescens), using the methods of construction of SSR-enrichment libraries and EST database mining of a related species P. fluviatilis. Of 16 genomic-SSR primer pairs examined, eight successfully amplified scorable products. The number of alleles at these informative loci varied from 3 - 14 with an average of 8.5 alleles per locus. When tested on wild perch from a population in Pennsylvania, observed and expected heterozygosities ranged from 0.07 - 0.81 and from 0.37 - 0.95, respectively. Of 2,226 EST sequences examined, only 110 (4.93%) contained microsatellites and for those, 13 markers were tested, 12 of which exhibited polymorphism. Compared with genomic-SSRs, EST-SSRs exhibited a lower level of genetic variability with the number of alleles of averaging only 2.6 alleles per locus. Cross-species utility indicated that three of the genomic-SSRs and eight of the EST-SSRs successfully cross-amplified in a related species, the walleye (Sander vitreus).
为了进行复杂性状的全基因组扫描分析,我们使用构建 SSR 富集文库和相关物种 P. fluviatilis 的 EST 数据库挖掘的方法,共分离和鉴定了 21 个新的基因组 SSR 和 EST-SSR,用于黄鲈(Perca flavescens)。在测试的 16 对基因组 SSR 引物中,有 8 对成功扩增出可评分的产物。这些信息性位点的等位基因数从 3-14 不等,平均每个位点 8.5 个等位基因。当在宾夕法尼亚州一个种群的野生黄鲈中进行测试时,观察到的和预期的杂合度分别在 0.07-0.81 和 0.37-0.95 之间。在测试的 2226 个 EST 序列中,只有 110 个(4.93%)含有微卫星,其中 13 个标记进行了测试,其中 12 个表现出多态性。与基因组 SSR 相比,EST-SSR 表现出较低的遗传变异性,每个位点的等位基因数平均只有 2.6 个。种间通用性表明,三个基因组 SSR 和八个 EST-SSR 成功地在相关物种大眼鲈(Sander vitreus)中扩增。