DYNAMICS OF MITOCHONDRIAL-DNA EVOLUTION IN ANIMALS - AMPLIFICATION AND SEQUENCING WITH CONSERVED PRIMERS
DYNAMICS OF MITOCHONDRIAL-DNA EVOLUTION IN ANIMALS - AMPLIFICATION AND SEQUENCING WITH CONSERVED PRIMERS
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DOI：
10.1073/pnas.86.16.6196
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发表时间：
1989-08-01
影响因子：
11.1
通讯作者：
WILSON, AC
中科院分区：
文献类型：
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作者：
KOCHER, TD;THOMAS, WK;WILSON, AC
通过一组针对保守区的标准引物，我们使用聚合酶链式反应从100多个动物物种中扩增出mtDNA的同源片段，包括哺乳动物、鸟类、两栖动物、鱼类和一些无脊椎动物。扩增和直接测序是可能的，使用来自纳克新鲜样本和微克量组织的未纯化线粒体DNA，这些组织在酒精中保存数月或在干燥状态下保存数十年。鸟类和鱼类的序列进化时带有与哺乳动物相同的强烈过渡倾向。然而，由于鸟类的轻链缺乏胸腺嘧啶，胸腺嘧啶到胞嘧啶的转变比其他类群中较少发生。哺乳动物和鸟类细胞色素b基因片段的氨基酸替换速度比鱼类快，替换模式符合细胞色素b的结构假说。这些引物出人意料的广泛分类用途为系统发育和种群研究提供了机会。
With a standard set of primers directed toward conserved regions, we have used the polymerase chain reaction to amplify homologous segments of mtDNA from more than 100 animal species, including mammals, birds, amphibians, fishes, and some invertebrates. Amplifications and direct sequencing were possible using unpurified mtDNA from nanogram samples of fresh specimens and microgram amounts of tissues preserved for months in alcohol or decades in the dry state. The bird and fish sequences evolve with the same strong bias toward transitions that holds for mammals. However, because the light strand of birds is deficient in thymine, thymine to cytosine transitions are less common than in other taxa. Amino acids replacement in a segment of the cytochrome b gene is faster in mammals and birds than in fishes and the pattern of replacements fits the structural hypothesis for cytochrome b. The unexpectedly wide taxonomic utility of these primers offers opportunities for phylogenetic and population research.