Dynamics of Mitochondrial DNA Evolution in Animals: Amplification and Sequencing with Conserved Primers

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. Amplification and direct sequencing were possi...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1989-08, Vol.86 (16), p.6196-6200
Main Authors: Kocher, T. D., Thomas, W. K., Meyer, A., Edwards, S. V., Pääbo, S., Villablanca, F. X., Wilson, A. C.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Amino acids
Animals
Base Sequence
Biological and medical sciences
Biological Evolution
Birds
Cytochrome b Group - genetics
Cytochromes
DNA
DNA, Mitochondrial - genetics
Evolution
Fundamental and applied biological sciences. Psychology
Gene Amplification
Genes
Genes. Genome
Mammals
Mitochondrial DNA
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
Protein Conformation
Rodents
Sequence Homology, Nucleic Acid
Sequencing
Species Specificity
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Summary: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. Amplification 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 acid 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.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.86.16.6196