Interspecific Recombination of Mitochondrial DNA Molecules in Hybrid Somatic Cells

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The mitochondrial DNA (mtDNA) in rodent-human hybrid somatic cells was studied in strains that contain nucleotide sequences from both parental mtDNAs. A test for linkage of rodent to human mtDNA was devised on the basis of the density and sequence differences between these DNAs. When a mixture of rodent and human mtDNAs was banded in a CsCl gradient and each fraction hybridized with a mixture of complementary [3H]RNA transcribed from human mtDNA and complementary [32P]RNA transcribed from rodent mtDNA, each DNA was detected as a distinct and separate band at its expected density by specific hybridization with its complementary RNA. In some of the hybrid cell strains, the mtDNA sequences derived from the two species did not separate in the CsCl gradients. This result is interpreted as evidence for linkage between sequences from the two parental mtDNAs. While the exact nature of the linkage and the structure of the molecules containing both types of sequences are not known, the evidence supports the conclusion that the linkage is realized by a covalent bond. An event leading to the covalent bonding of these different sequences may be described as recombination. Among 18 hybrid cell strains examined, 13 contained large proportions of recombinant molecules. These molecules were found in both mouse-human and rat-human strains, and in strains containing more human or more rodent mtDNA sequences.

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