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Mitochondrial DNA genotypes in nuclear transfer-derived cloned sheep

Abstract

Eukaryotic cells contain two distinct genomes. One is located in the nucleus (nDNA) and is transmitted in a mendelian fashion, whereas the other is located in mitochondria (mtDNA) and is transmitted by maternal inheritance. Cloning of mammals1,2,3,4,5,6 typically has been achieved via nuclear transfer, in which a donor somatic cell is fused by electoporation with a recipient enucleated oocyte. During this whole-cell electrofusion, nDNA as well as mtDNA ought to be transferred to the oocyte7,8. Thus, the cloned progeny should harbour mtDNAs from both the donor and recipient cytoplasms, resulting in heteroplasmy. Although the confirmation of nuclear transfer has been established using somatic cell-specific nDNA markers, no similar analysis of the mtDNA genotype has been reported. We report here the origin of the mtDNA in Dolly, the first animal cloned from an established adult somatic cell line, and in nine other nuclear transfer-derived sheep generated from fetal cells. The mtDNA of each of the ten nuclear-transfer sheep was derived exclusively from recipient enucleated oocytes, with no detectable contribution from the respective somatic donor cells. Thus, although these ten sheep are authentic nuclear clones, they are in fact genetic chimaeras, containing somatic cell-derived nuclear DNA but oocyte-derived mtDNA.

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Figure 1: The sheep D-loop region.
Figure 2: PCR/RFLP analyses of the 544-bp PCR-amplified D-loop fragments, showing the maps with the predicted digestion fragment sizes, in bp, for each of the 4 indicated restriction endonucleases.

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Acknowledgements

We thank E.A. Shoubridge, M. Hirano and S.C. Silverstein for helpful discussions. This work was supported by grants from the National Institutes of Health (NS28828; NS11766; HD32062; HL52145) and the Muscular Dystrophy Association.

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Correspondence to Eric A. Schon.

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Evans, M., Gurer, C., Loike, J. et al. Mitochondrial DNA genotypes in nuclear transfer-derived cloned sheep. Nat Genet 23, 90–93 (1999). https://doi.org/10.1038/12696

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  • DOI: https://doi.org/10.1038/12696

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