FitzSimmons N N, Moritz C, Moore S S
Department of Zoology, University of Queensland, Brisbane, Australia.
Mol Biol Evol. 1995 May;12(3):432-40. doi: 10.1093/oxfordjournals.molbev.a040218.
Microsatellite loci consisting of (CA)n repetitive arrays were obtained from three species of marine turtle, and primers were designed to test for polymorphism within species and the persistence of microsatellites across species. Homologous loci were found in each test of six marine species within two families (Cheloniidae and Dermochelyidae), as well as in a freshwater species (Emydidae, Trachemys scripta), which indicates a conservation of flanking sequences spanning approximately 300 million years of divergent evolution. The persistence of homologous microsatellites across marine turtles was confirmed by direct sequencing of loci across species and by the discovery of polymorphism in 24 of 30 cross species tests. The conservation of flanking sequences could be due to a slow rate of base substitution in turtle nuclear DNA, as previously reported for mtDNA. In contrast, the presence of up to 25 alleles per locus per species indicates that the replication slippage events responsible for changes in allele length operate as in mammals. Comparisons of alleles among species revealed that alleles of the same length may not be homologous due to mutations within the flanking sequences. Levels of heterozygosity were consistently higher in species from which the primers were designed, which suggests problems with cross-species comparisons of variability. Within species, microsatellite variation between divergent populations was consistent with results from previous mtDNA studies indicating the usefulness of microsatellites for comparing male- versus female-mediated gene flow.
从三种海龟中获取了由(CA)n重复序列组成的微卫星位点,并设计了引物来检测种内多态性以及微卫星在不同物种间的持久性。在两个科(海龟科和棱皮龟科)的六种海洋物种以及一种淡水物种(泽龟科,滑龟)的每次测试中都发现了同源位点,这表明侧翼序列在约3亿年的分歧进化过程中得以保守。通过对跨物种位点的直接测序以及在30次跨物种测试中的24次发现多态性,证实了同源微卫星在海龟中的持久性。侧翼序列的保守性可能是由于海龟核DNA中的碱基替换速率较慢,正如之前对线粒体DNA的报道。相比之下,每个物种每个位点最多有25个等位基因,这表明导致等位基因长度变化的复制滑动事件与哺乳动物中的情况相同。物种间等位基因的比较显示,由于侧翼序列内的突变,相同长度的等位基因可能并非同源。设计引物的物种的杂合度水平一直较高,这表明在跨物种变异性比较方面存在问题。在物种内部,不同种群间的微卫星变异与之前线粒体DNA研究的结果一致,表明微卫星在比较雄性和雌性介导的基因流方面具有实用性。