Mitochondrial DNA genotypes in nuclear transfer-derived cloned sheep.
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 mammals 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 oocyte. 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.
Evans
M J
MJ
Integrated Program in Cellular, Molecular and Biophysical Studies, Columbia University College of Physicians and Surgeons, 630 West 168th Street, New York, New York 10032, USA.
Gurer
C
C
Loike
J D
JD
Wilmut
I
I
Schnieke
A E
AE
Schon
E A
EA
D000818:Animals; D001483:Base Sequence; D002467:Cell Nucleus / Q000235:genetics; D002678:Chimera; D019976:Cloning, Organism; D004272:DNA, Mitochondrial; D005347:Fibroblasts; D005838:Genotype; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000378:metabolism; D009154:Mutation; D009865:Oocytes / Q000378:metabolism; D010920:Placenta / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D012756:Sheep / Q000235:genetics*
D004272:DNA, Mitochondrial