Figure 1.
The Right Back Leg of the Woolly Mammoth(M. primigenius) Found in Siberia
The well-preserved mammoth body fragment with foot (33 × 36 cm), shin, and ankle-joint (the total length is ~88 cm) was found in the Enmynveem River valley (north-eastern Siberia, Chukotka). The tissue material (bones, muscles, and skin) had no visible marks of tissue damage by insects or other animals. Radiocarbon dating of the skin and muscle tissue determined that the mammoth lived 32,850 ± 900 y ago [12].
Figure 2.
The Unusually Well-Preserved Mammoth DNA
(A) Nuclei with DNA clearly detectable by DAPI staining in muscle cells of ~33,000-y-oldM. primigenius. (B) Total genomic DNA isolated from the mammoth muscle tissue (lane 1 is 1/10 dilution of the DNA on lane 2); control DNA from fresh human blood samples in lanes 3 and 4. (C) Examples of PCR products (~300–600 bp) for mammoth mitochondrial genome. (D) PCR amplification recovers long sequences for complete mitochondrial genes (1,317 bpCytB and 1,613 bpATP6 genes), but PCR of larger fragments (3,054 bpND5) is failed.
Figure 3.
Mitochondrial Genome of the Woolly MammothM. primigenius
The complete mitochondrial genome was determined independently in two different laboratories using designs with multiple primers for overlapping PCR fragments ranged from ~325 to ~650 bp; the longer PCR fragments were also produced and sequenced. The overlapped PCR products used for sequencing and cloning are shown by the inner circle. Only PCR fragments produced from different pairs of primers are shown. Two genes,ATP6 andND4L, overlap with neighboring genes.
Figure 4.
Paenungulata Tree and Phylogenic Relationship of the Woolly Mammoth
The analysis of complete mtDNA sequences placesM. primigenius withE. maximus on the tree. The Sirenia(D. dugon) and Hyracoidea(P. capensis), most closely related species among extant taxa to Elephantinae, were taken as outgroups. Bootstrap values and posterior probabilities were calculated using a Bayesian approach [29,31] assuming a gamma distribution of the rates of evolution across sites with a General Time Reversible model (normal font), HKY model (bold), with a parsimony approach (italic), and by neighbor joining (italic and bold) [28]. The scale is 0.1 substitutions per site. The mitochondrial genomes ofM. primigenius, E. maximus A, E. maximus B, L. africana A, L. africana B, D. dugon, andP. capensis were used in the analysis.